Phage display technology
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Theoretically, phage display is an exogenous gene expression method which the gene encoding the interest protein is inserted into bacteriophage coat protein gene then displaying the interest protein on the phage surfaces, resulting in a connection between genotype and phenotype.https://www.creative-biolabs.com/phage-display-service.html
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![Phage
Display
Technology
in Creative
Biolabs
Creative Biolabs is a leading service provider of phage display library construction and screening. In a phage
display library, a variety of peptides, small antibodies [e.g. scFv and Fab] or proteins are displayed on the
surface of filamentous phage [M13, fd, and f1 strains] as fusion proteins with one of the coat proteins of the
phage virions, while the genetic materials encoding the peptides/proteins are housed within the virions. Using a
binding-based process called biopanning, a small number of phages that display proteins/peptides specifically
binding to a target of interest can be rescued from a phage library that usually displays a repertoire of many
billions of unique peptides/proteins. Finally, the peptides/proteins displayed by the selected phages can be
identified by phage amplification and DNA sequencing.
Phage Display Technology
Phage Display Systems in Creative Biolabs
M13 Phage Display
pVIII-fusion display: major coat protein fusion, display up to 2700 copies of foreign protein
pIII-fusion display: minor protein, display 1-5 copies of foreign protein
Most popular option for phage display, has been applied in many different research areas.
T4 Phage Display
Larger genome DNA which enables larger insertions
Dual display: two different molecules can be displayed separately on HOC and SOC
Both N- and C-terminal insertion available
T7 Phage Display
Time saving: T7 phages have a shorter lifecycle than filamentous phages and lambda phages.
Optimized biopanning: as T7 phages are resistant to extreme conditions, a variety of agents can
be applied in screening procedure in contrast to alternative phages.
Complementary to M13 phage, widely used for cDNA library.
Library Construction Service in Creative Biolabs
Phage Display Library Types
Antibody library (immune, naïve, semi-synthetic, synthetic)
Peptide library (linear, cyclic)
Protein scaffold library
cDNA Library
Mutagenesis Strategies for Library Construction
Trimer codon method
Mutations are introduced at the codon level rather than at individual bases
No codon bias, no frame shift, no stop codon
Defined AA composition at each position
Kunkel-like oligonucleotide-directed mutagenesis method
Degenerate codon method
Features
No system-based out-of-frame mutations
No limit on target length
Flexible and scalable
Large library capacity: from 108 to over 1011
Premade Antibody Libraries Available!
During the past decades, we have integrated a comprehensive portfolio of premade libraries,
including linear and cyclic peptide libraries, monobody libraries human scFv libraries, human
Fab libraries, mouse scFv/Fab libraries, rabbit Fab libraries, chicken scFv libraries and single
domain antibody libraries.
Library Screening Service in Creative Biolabs
Tailored Biopanning Strategies:
Solid-phase screening
Phage libraries are selected by flowing through a solid surface with the immobilized target.
In-solution screening
Isolating binders recognize naïve targets. The target-binder interaction is carried out in solution
with subsequent capture by the appropriate method.
Cell-based screening
It is suitable to select peptides/antibodies for cell surface receptors, such as GPCR and ion
channel-linked receptor.
In Vivo screening
Isolating novel peptides as the functional markers of new receptors or novel drug target
candidates.
Applications of Phage Display Technology
Creative Biolabs offers high-quality phage display library construction and custom phage display
library screening services for a broad range of project objectives including but not limited to:
Function peptide discovery
Therapeutic antibody discovery
Monoclonal antibody discovery from a variety of species includes: human, monkey, llama,
camel, shark, alligator, mouse, rat, hamster, guinea pig, rabbit, chicken, dog, bovine, goat,
sheep, and ferret.
Antibody humanization
Antibody affinity maturation
Contact Us
Email: info@creative-biolabs.com
Web: www.creative-biolabs.com
Tel: 1-631-381-2994 Fax: 1-631-207-8356
Address: 45-1 Ramsey Road, Shirley, NY 11967, USA](https://image.slidesharecdn.com/phagedisplaytechnologyflyer-210408090805/85/Phage-display-technology-1-320.jpg)
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Phage display and its applications
This document discusses bacteriophages and their use in phage display. Specifically, it notes that bacteriophages infect bacterial cells and use them to replicate viruses. It then explains that phage display involves fusing foreign genes or proteins to the surface of phages, creating libraries of phages that each display a single protein. These libraries can be exposed to targets, and phages that interact are selected and amplified through multiple rounds. The document outlines several applications of proteins isolated through phage display, such as epitope mapping, drug discovery, and developing new vaccines or treatments that have a specific interaction with a target antigen, protein, or disease.
Site directed mutagenesis by pcr
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DNA libraries
Genomic DNA libraries contain representative copies of all DNA fragments in an organism's genome, including both expressed and non-expressed sequences. They are constructed by isolating genomic DNA, fragmenting it, and cloning the fragments into suitable vectors like lambda phage or BACs. cDNA libraries contain only expressed sequences, as they are constructed by isolating mRNA from tissues, reverse transcribing it to cDNA, and cloning the cDNA fragments. Both library types are useful for gene discovery, sequencing, mapping genomes, and studying regulatory sequences.
Phage display
Phage display technology allows the display of proteins or peptides on the outside of bacteriophages while encoding the corresponding gene on the inside. This allows for large libraries to be screened in vitro to select for interactions between the displayed molecules and a target. The most common phages used are filamentous phages like M13, which can be genetically manipulated to display proteins of interest. The technique involves inserting a gene into the phage coat protein gene, infecting bacteria to produce phage particles displaying the protein, and panning against a target to isolate interacting proteins.
Dna library lecture-Gene libraries and screening
This document discusses gene libraries and screening procedures. It begins by explaining what genomic and cDNA libraries are. It then provides details on creating genomic libraries, including purifying genomic DNA, fragmenting it, and cloning the fragments into vectors. Creating cDNA libraries involves isolating mRNA, synthesizing cDNA, and ligating the cDNA to vectors. The size of libraries needed to ensure coverage of genomes is calculated. Lambda phage is described as a commonly used vector that can accept inserts up to 23kb in size. The processes of packaging recombinant DNA into lambda phage particles and creating lambda phage libraries are outlined.
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Proteoimic presentation
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Protein protein interactions
The document discusses protein-protein interactions (PPIs) and methods used to study them. It defines PPIs as physical contacts between two or more proteins through biochemical or electrostatic forces. It describes different types of PPIs including homo-oligomers, hetero-oligomers, covalent and non-covalent interactions. Common methods to study PPIs are also summarized, such as yeast two-hybrid systems, co-immunoprecipitation, and protein interaction databases. The applications and importance of PPI research are mentioned including roles in various cellular processes and diseases.
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Phage display and its applications
This document discusses bacteriophages and their use in phage display. Specifically, it notes that bacteriophages infect bacterial cells and use them to replicate viruses. It then explains that phage display involves fusing foreign genes or proteins to the surface of phages, creating libraries of phages that each display a single protein. These libraries can be exposed to targets, and phages that interact are selected and amplified through multiple rounds. The document outlines several applications of proteins isolated through phage display, such as epitope mapping, drug discovery, and developing new vaccines or treatments that have a specific interaction with a target antigen, protein, or disease.
Site directed mutagenesis by pcr
Site-directed mutagenesis is a molecular biology technique used to make specific changes to DNA sequences. It involves using a primer containing the desired mutation in a PCR reaction to introduce the mutation into the gene of interest. There are different approaches for site-directed mutagenesis using PCR, including using a mutated primer in normal PCR or a primer extension method. The technique is used for applications like protein engineering to study the impact of sequence changes or insert restriction sites. However, it can be difficult to replicate the mutated DNA and screening mutations requires sequencing.
DNA libraries
Genomic DNA libraries contain representative copies of all DNA fragments in an organism's genome, including both expressed and non-expressed sequences. They are constructed by isolating genomic DNA, fragmenting it, and cloning the fragments into suitable vectors like lambda phage or BACs. cDNA libraries contain only expressed sequences, as they are constructed by isolating mRNA from tissues, reverse transcribing it to cDNA, and cloning the cDNA fragments. Both library types are useful for gene discovery, sequencing, mapping genomes, and studying regulatory sequences.
Phage display
Phage display technology allows the display of proteins or peptides on the outside of bacteriophages while encoding the corresponding gene on the inside. This allows for large libraries to be screened in vitro to select for interactions between the displayed molecules and a target. The most common phages used are filamentous phages like M13, which can be genetically manipulated to display proteins of interest. The technique involves inserting a gene into the phage coat protein gene, infecting bacteria to produce phage particles displaying the protein, and panning against a target to isolate interacting proteins.
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This document discusses gene libraries and screening procedures. It begins by explaining what genomic and cDNA libraries are. It then provides details on creating genomic libraries, including purifying genomic DNA, fragmenting it, and cloning the fragments into vectors. Creating cDNA libraries involves isolating mRNA, synthesizing cDNA, and ligating the cDNA to vectors. The size of libraries needed to ensure coverage of genomes is calculated. Lambda phage is described as a commonly used vector that can accept inserts up to 23kb in size. The processes of packaging recombinant DNA into lambda phage particles and creating lambda phage libraries are outlined.
P bluescript
pBluescript is an example of a combination between plasmids and phages (phagemids).
Phagemids represent a hybrid type of class of vectors that serve to produce single-stranded DNA.
Proteoimic presentation
Protein-protein interactions (PPIs) are important for many cellular functions. There are two main types of PPIs - transient interactions which are brief, and stable interactions which form multiprotein complexes. Crosslinking can capture both transient and stable PPIs by covalently binding interacting proteins. In vivo crosslinking studies PPIs in their native environment while in vitro crosslinking allows better reaction control. Pull-down assays use affinity purification to isolate stable protein complexes and identify binding partners of a bait protein. SDS-PAGE is commonly used to separate and visualize proteins isolated by techniques like pull-down.
Protein protein interactions
The document discusses protein-protein interactions (PPIs) and methods used to study them. It defines PPIs as physical contacts between two or more proteins through biochemical or electrostatic forces. It describes different types of PPIs including homo-oligomers, hetero-oligomers, covalent and non-covalent interactions. Common methods to study PPIs are also summarized, such as yeast two-hybrid systems, co-immunoprecipitation, and protein interaction databases. The applications and importance of PPI research are mentioned including roles in various cellular processes and diseases.
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2. Total RNA is first extracted using Trizol, which separates RNA, DNA and proteins. mRNA is then isolated from total RNA using biotin-labeled oligo dT probes that selectively bind to the poly-A tail of mRNA molecules.
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Proteome analysis
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Selectable markers
Thymidine kinase (TK), dihydrofolate reductase (dhfr), and chloramphenicol acetyl transferase (CAT) are examples of selectable marker genes used for animal cells. Marker genes help monitor transfection by allowing detection of whether the transgene was successfully transferred. Selectable marker genes enable transformed cells to survive selection conditions that kill non-transformed cells.
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MPSS is a technique for analyzing gene expression that involves sequencing cDNA fragments cloned onto microbeads. It allows for the simultaneous sequencing of over 1 million cDNA clones. MPSS generates 17-base signature sequences that uniquely identify mRNA transcripts. Gene expression levels are quantified by counting the number of signatures for each gene. MPSS provides a more in-depth analysis of gene expression compared to other methods as it can detect genes expressed at very low levels and does not require prior knowledge of gene sequences.
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The document discusses the process of synthesizing cDNA from mRNA. It involves isolating mRNA, using reverse transcriptase to copy the mRNA into single-stranded cDNA, then converting it to double-stranded cDNA using DNA polymerase. The double-stranded cDNA can then be inserted into a vector and used to create a cDNA library through cloning in bacteria or phage. The library can be screened by hybridization or assays to identify clones containing genes of interest.
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Automated sequencing of genomes require automated gene assignment
Includes detection of open reading frames (ORFs)
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Phage panning
Library Construction and Screening
Fig.1 Workflow of phage display construction
and screening (Díez et al. 2015)
Phage display is one of the most powerful and widely used laboratory technique for the study of protein-protein, protein-peptide and protein-DNA interactions. This technology is mainly based on displaying the interest protein (peptides, antibodies, scaffolds or others) on the surface of employing phage and then be used to interrogate the constructed libraries containing millions or even billions of displayed phages. Theoretically, phage display is an exogenous gene expression method which the gene encoding the interest protein is inserted into bacteriophage coat protein gene then displaying the interest protein on the phage surfaces, resulting in a connection between genotype and phenotype.
Phage panning
Creative Biolabs has long-term devoted to the development and application of phage display technology. With years of experience, our scientists have developed several phage display based platforms and tailored hundreds of particular libraries and thousands of specific antibody products to boost our global customers’ research and project goals. We are pleased to use our extensive experience and advanced platform to offer the best service and the most qualified products to satisfy each demand from our customers.
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shotgun sequncing
Genome sequencing is the process of determining the order of nucleotide bases - A, C, G, and T - that make up an organism's DNA. Shotgun sequencing involves randomly breaking the genome into small fragments, sequencing those pieces, and reassembling the sequence by identifying overlapping regions. It was originally used by Sanger to sequence small genomes like viruses and bacteria. There are two main methods - hierarchical shotgun sequencing for larger genomes containing repeats, and whole genome shotgun sequencing for smaller genomes.
Sage
SAGE is a technique that allows for the digital analysis of overall gene expression patterns through the use of short sequence tags to uniquely identify transcripts without requiring preexisting clones. It works by linking these tags together into long serial molecules that can then be cloned and sequenced, with the number of times a particular tag is observed providing the expression level of the corresponding transcript. This allows for rapid sequencing analysis of multiple transcripts from a single sequencing event. SAGE is useful for comparative expression studies to identify differences in gene expression between tissues.
Roche Pyrosequencing 454 ; Next generation DNA Sequencing
In these slides we are describing all about the DNA sequencing technique which will based on pyrosequencing termed as roche 454 pyrosequencing.
Pyrosequencing
Pyrosequencing is a sequencing method that detects DNA polymerase activity by measuring the release of pyrophosphate using a cascade of enzymatic reactions that generate visible light. It utilizes emulsion PCR to amplify DNA fragments on beads in microreactors. The beads are then loaded into wells and sequenced by sequentially adding nucleotides and detecting light produced upon incorporation using a CCD camera. Key advantages are its accuracy, high throughput of up to 48,000 probes per day, and ease of automation. However, it requires specialized equipment and software.
cohesive and blunt end ligation
This document discusses various methods for ligating DNA fragments, including blunt end ligation, sticky end ligation using linkers or adaptors, and homopolymeric tailing. Blunt end ligation is less efficient than sticky end ligation. Linkers and adaptors are oligonucleotides used to create sticky ends for ligation, while homopolymeric tailing uses terminal transferase to add homopolymer tails to blunt ends before ligation. The goal is to efficiently join vector and insert DNA fragments for recombinant DNA construction.
Preparation of competent cells
This document provides instructions for preparing competent bacterial cells for transformation. Key steps include growing cells to mid-log phase, treating with calcium chloride to make membranes permeable, adding exogenous DNA, and applying a heat shock to induce DNA uptake. The resulting transformed competent cells can be used to take up exogenous DNA from their surroundings through their membranes. Proper preparation is important for achieving high transformation efficiency.
mRNA Isolation
1. mRNA isolation is the process of extracting messenger RNA from biological samples. It is important for research and industry applications as mRNA provides insight into which genes are being expressed and translated into proteins.
2. Total RNA is first extracted using Trizol, which separates RNA, DNA and proteins. mRNA is then isolated from total RNA using biotin-labeled oligo dT probes that selectively bind to the poly-A tail of mRNA molecules.
3. The mRNA-probe complexes are immobilized on magnetic beads coated with streptavidin. This allows the mRNA to be separated and purified from other RNAs through magnetic separation washes. The purified mRNA can then be used in applications like RT-PCR and protein
sequence of file formats in bioinformatics
This document discusses different biological database file formats for storing sequence and molecular data. It describes several common formats including FASTA, Multi-FASTA, GeneBank flat-file format, GCG, EMBL, Clustal, and SWISS PROT. It provides details on the header, feature, and sequence sections of each format and how they represent genetic and protein information.
Chromosome walking
Chromosome walking is a method used to locate and clone a specific gene or allele through successive identification of overlapping DNA sequences. It begins at a known marker gene near the target and "walks" through testing genes one by one to map their locations and identify overlaps, eventually reaching the mutant gene. Once the full sequence is cloned, the gene's function can be determined to study genetically transmitted diseases. It is a complex process but has allowed mapping of large chromosome regions over 1000kb.
2d Page
2D-PAGE is a technique used to separate complex protein mixtures based on isoelectric point and molecular weight. It involves two sequential steps - isoelectric focusing and SDS-PAGE. In isoelectric focusing, proteins are separated based on their isoelectric point in an immobilized pH gradient. They are then separated by SDS-PAGE based on their molecular weight. The separated proteins can then be visualized through staining and identified through mass spectrometry. While useful for proteomic analysis, 2D-PAGE has limitations such as low reproducibility and dynamic range.
ZINC FINGER NUCLEASE TECHNOLOGY
Zinc finger nucleases (ZFNs) allow for highly targeted editing of the genome. ZFNs consist of a DNA-binding domain made of zinc finger proteins and a DNA-cleaving domain. The ZFN pair binds to a target site and creates a double-strand break, which the cell repairs through non-homologous end joining or homologous recombination, enabling gene knockouts or targeted changes. ZFNs work in many cell types and animal models, providing a more efficient alternative to traditional transgenic techniques. They have applications in functional genomics, cell line engineering, and animal model generation.
Viral vector
This document discusses viral vectors, which are tools used by molecular biologists to deliver genetic material into cells. It describes how viruses are efficient at transferring their DNA into host cells and can be modified to insert exogenous genes. The document outlines some key properties of viral vectors like safety through deletion of viral replication genes. It then describes some common viral vectors, including adenovirus, adeno-associated virus, herpes virus, lentivirus, and retrovirus. Finally, it discusses some applications of viral vectors like gene therapy and using viruses expressing pathogen proteins as vaccines.
Proteome analysis
Proteome analysis studies the array of proteins expressed in a biological system under particular conditions. It can help understand cellular pathways and biological processes by characterizing protein complexes. Proteome analysis is also used to discover disease biomarkers for diagnostics and drug development by identifying protein expression changes. Key developments driving the field include improvements to 2D electrophoresis for separating proteins, mass spectrometry for analyzing separated proteins, and bioinformatics tools for searching protein databases. Common steps in proteome analysis involve separating proteins, digesting them into peptides, and identifying peptides and proteins using mass spectrometry techniques like MALDI-TOF-MS and ESI-Q-IT-MS.
Selectable markers
Thymidine kinase (TK), dihydrofolate reductase (dhfr), and chloramphenicol acetyl transferase (CAT) are examples of selectable marker genes used for animal cells. Marker genes help monitor transfection by allowing detection of whether the transgene was successfully transferred. Selectable marker genes enable transformed cells to survive selection conditions that kill non-transformed cells.
Ion torrent
The document summarizes Ion Torrent sequencing technology. It detects hydrogen ions released during DNA polymerization rather than using optics. The sequencing occurs on semiconductor chips patterned through photolithography into wells, each sequencing a different template. As nucleotides are incorporated, hydrogen ions change the pH detected by ion sensors below each well. This allows massively parallel sequencing that is faster, cheaper and simpler than previous technologies.
MASSIVELY PARELLEL SIGNATURE SEQUENCING
MPSS is a technique for analyzing gene expression that involves sequencing cDNA fragments cloned onto microbeads. It allows for the simultaneous sequencing of over 1 million cDNA clones. MPSS generates 17-base signature sequences that uniquely identify mRNA transcripts. Gene expression levels are quantified by counting the number of signatures for each gene. MPSS provides a more in-depth analysis of gene expression compared to other methods as it can detect genes expressed at very low levels and does not require prior knowledge of gene sequences.
Tumor formtion , ti ri plasmid , dna trnsfr.
This document summarizes information about tumor formation in plants caused by Agrobacterium tumefaciens and Agrobacterium rhizogenes bacteria. It discusses how the Ti and Ri plasmids are transferred into plant cells, causing crown gall and hairy root diseases respectively. The Ti plasmid contains T-DNA which is integrated into the plant genome, inducing tumor formation and opine synthesis. DNA transfer techniques like electroporation, microprojectile bombardment, and microinjection are also summarized for introducing foreign genes into plant cells.
Synthesis of c dna
The document discusses the process of synthesizing cDNA from mRNA. It involves isolating mRNA, using reverse transcriptase to copy the mRNA into single-stranded cDNA, then converting it to double-stranded cDNA using DNA polymerase. The double-stranded cDNA can then be inserted into a vector and used to create a cDNA library through cloning in bacteria or phage. The library can be screened by hybridization or assays to identify clones containing genes of interest.
Gene prediction methods vijay
Automated sequencing of genomes require automated gene assignment
Includes detection of open reading frames (ORFs)
Identification of the introns and exons
Gene prediction a very difficult problem in pattern recognition
Coding regions generally do not have conserved sequences
Much progress made with prokaryotic gene prediction
Eukaryotic genes more difficult to predict correctly
Binary Vector, By KK Sahu sir
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
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and screening (Díez et al. 2015)
Phage display is one of the most powerful and widely used laboratory technique for the study of protein-protein, protein-peptide and protein-DNA interactions. This technology is mainly based on displaying the interest protein (peptides, antibodies, scaffolds or others) on the surface of employing phage and then be used to interrogate the constructed libraries containing millions or even billions of displayed phages. Theoretically, phage display is an exogenous gene expression method which the gene encoding the interest protein is inserted into bacteriophage coat protein gene then displaying the interest protein on the phage surfaces, resulting in a connection between genotype and phenotype.
Phage panning
Creative Biolabs has long-term devoted to the development and application of phage display technology. With years of experience, our scientists have developed several phage display based platforms and tailored hundreds of particular libraries and thousands of specific antibody products to boost our global customers’ research and project goals. We are pleased to use our extensive experience and advanced platform to offer the best service and the most qualified products to satisfy each demand from our customers.
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https://www.creative-biolabs.com/sdab/sdab-functional-identification.htm
Sdab discovery
Single Domain Antibody also known as domain antibody, VHH, VNARor sdAb, is a kind of antibody fragments consisting of a single monomeric variable antibody domain and lacking the light chain and CH domain of the heavy chain in conventional Fab region.https://www.creative-biolabs.com/sdab/one-stop-solution-for-sdab-development.htm
Library screening
This document discusses different types of DNA libraries and methods for screening libraries to identify clones containing genes of interest. It describes genomic and cDNA libraries, noting that genomic libraries contain all DNA fragments from an organism's genome while cDNA libraries contain only coding sequences. The key screening methods discussed are colony/plaque hybridization using radiolabeled probes, expression screening using antibodies, and PCR screening using gene-specific primers.
Antibody mouse
Creative Biolabs is an undisputed world-leading service provider and research institution in phage display library technique. We offer following world-class premade human and mouse antibody libraries with great capacity & diversity that can derive highly specific, high affinity antibodies ranging from 10 pM to 10 nM.
Human antibody library
Creative Biolabs is an undisputed world-leading service provider and research institution in phage display library technique. We offer following world-class premade human and mouse antibody libraries with great capacity & diversity that can derive highly specific, high affinity antibodies ranging from 10 pM to 10 nM.
Genomic library
Genomic libraries are collections of bacteria that have been engineered to contain the entire DNA of an organism. DNA from the organism is cut into fragments and inserted into cloning vectors like plasmids within bacteria. Together the bacteria hold the complete genome. Genomic libraries allow researchers to isolate and study specific DNA sequences using probes, map genomes, and clone DNA segments for various research purposes like genetic modification of crops. They are constructed by extracting, cutting, and ligating an organism's DNA into vectors within host bacteria. Different vectors have varying capacities to hold DNA fragments, ranging from plasmids at 15kb to yeast artificial chromosomes at up to 2,000kb.
Affinity maturation of antibodies
Subsequent library screening will fish out the antibody mutants that have high affinity. Two library screening strategies are available. In the first "surface-panning" strategy, decreasing concentrations of antigen is surface immobilized. In the second "solution-sorting" strategy, in which a labeled antigen in solution is used, we have two approaches, selection based on the equilibrium constant (Kd) and selection based on binding kinetics. In the first approach, sub-library phage is incubated with biotinylated antigen at controlled concentrations and bound phages are captured by immobilized NeutrAvidin. Selection based on binding kinetics is also termed off-rate (Koff) selection, in which phage population is allowed to saturate the labeled antigen before a large molar excess of unlabeled antigen is added to the mix for controlled periods of time. This allows the selection of mutant antibodies that have slower off-rates. Since a reduction in Koff usually results in a higher affinity, this selection approach singles out antibody variants with improved Kd.
Antibody library screening
Antibodies can be isolated
from recombinant antibody
libraries in lab. Using one of
these platforms for selection
that in essence mimics in vivo
process.
Antibody library screening
Antibodies can be isolated from recombinant antibody libraries in lab. Using one of these platforms for selection that in essence mimics in vivo process.https://www.creative-biolabs.com/phage-display-library-screening.html
Magic phage
As alternatives, Creative Biolabs also offers transgenic mouse/rat hybridoma and Native plasma cell sorting strategies which can also incorporate with transgenic mice for antibody development. Our experienced scientists are always willing to discuss with every customer on a case-to-case basis. Please feel free to inquiry us for more details.
DNA Library
Genomic and cDNA libraries allow for the representation of genomic sequences as multiple small fragments. A genomic library contains fragments from all DNA sources, including coding and non-coding regions. A cDNA library contains only expressed coding sequences, as it is synthesized from mRNA. The process involves isolating mRNA, synthesizing cDNA, incorporating it into a vector, and cloning the fragments. Libraries are important tools for studying genomes, genes, and gene expression.
Antibody avidity
We use an error-prone PCR integrated DNA-shuffling approach to mutate mainly CDR regions during sub-library construction. If the potential of introducing immunogenic mutations to framework positions is not a concern, we usually use this approach to create mutations at completely random positions across the entire VH and VL fragments. In these cases, the genetic diversity of the sub-library is further increased via passage through our proprietary bacterial mutator strain, CD-affi™.
PROTEIN MICROARRAYS
it will help you to understand how the protein microarrays are made, what are the different types and what all purposes they are used for. its very useful ppt
protein microarray-types and approaches.pptx
Protein microarrays are a high-throughput technology that allows thousands of proteins to be analyzed simultaneously. They consist of a solid support coated with thousands of different proteins in a defined array. Each protein spot represents a different protein. Protein microarrays can be used to study protein expression levels, interactions, activities, and functions on a large scale. There are different types including analytical, functional, and reverse-phase microarrays that each have their own strengths and applications.
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One-Stop Antibody Drug Discovery Services from GenScript ProBio
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Phage Display Screening for Single Domain Antibody (sdAb)
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ADC conjugate sites analysis
Antibody-drug conjugates employ the specific monoclonal antibodies (mAbs) to achieve targeted delivery of the conjugated cytotoxic molecules to tumor cells.
https://www.creative-biolabs.com/adc/conjugate-sites-analysis.htm
adc chemistry
As one major component of an antibody-drug conjugate (ADC), the antibody is the key for target specificity and serves as the cargo to deliver the cytotoxic drug (payload).
https://www.creative-biolabs.com/adc/antibody-design-and-conjugation.htm
ADC Biotechnology
To retain antibody bioactivity, mild, near-physiological conditions are often used for conjugation reactions. Under these conditions, endogenous amino acids such as Lys and Cys are chemically reactive and can be used as conjugation sites.https://www.creative-biolabs.com/adc/antibody-design-and-conjugation.htm
Antibody Design and Conjugation
As one major component of an antibody-drug conjugate (ADC), the antibody is the key for target specificity and serves as the cargo to deliver the cytotoxic drug (payload). A payload drug can be attached to different sites on an antibody using diverse conjugation chemistry. Multiple endogenous amino acids can serve as potential conjugation sites. However, to achieve more precisely controlled site-directed conjugations and subsequently a narrower distribution of drug-to-antibody ratio (DAR), special moieties with unique conjugation chemistries are engineered into antibody sequences in our antibody design services.
https://www.creative-biolabs.com/adc/antibody-design-and-conjugation.htm
adc cd40
CD40, also known as TNFRSF5, is a type I transmembrane protein. The molecular weight of CD40 is 48-kDa and it consists of a 193 amino acid (aa) extracellular domain, 21 aa leader sequence, 22 aa transmembrane domain, and a 62 aa intracellular domain in human (90 aa in mouse).https://www.creative-biolabs.com/adc/target-cd40-122.htm
ADC Biochemical Analysis
ADC preparation involves the chemical conjugation of the three components and depending on the conjugation strategy used, this process often yields complex and heterogeneous products.https://www.creative-biolabs.com/adc/adc-biochemical-analysis.htm
ADC Antibody Screening
While conventional cancer therapies (surgery, chemo therapy, and radiation therapy) have shown some success in the battle again cancer, they are often accompanied by complex and sometimes, severe side-effects due to the lack of target specificity. To circumvent this flaw and improve the efficacy and safety of cancer treatment, targeted cancer therapies, especially antibody-drug conjugates (ADCs), have been actively exploited and they are gaining a significant amount of attention during the recent years.https://www.creative-biolabs.com/adc/adc-antibody-screening.htm
ADC Affinity Measurement
The elegant design of an antibody-drug conjugate is designated to achieve targeted delivery of the conjugated cytotoxic agents to tumor cells and drug release upon antigen binding and internalization, thus maximizing the antitumor effects while minimizing cytotoxicity to normal tissues. The efficacy of an ADC greatly depends on the specific antigen binding activities of the monoclonal antibody (mAb) portion of the molecule.https://www.creative-biolabs.com/adc/adc-affinity-measurement.htm
5t4 adc
Development of 5T4-based Bispecific ADCs
A bispecific antibody can bind two different targets or two distinct epitopes on the same target. 5T4, specifically overexpressed on the cell surface of various tumors and internalized rapidly when bound to antibody, may be used as an attractive target to develop effective immunotherapy such as bispecific antibody-drug conjugate (ADC).
https://www.creative-biolabs.com/adc/development-of-5t4-based-bispecific-adcs.htm
IND Publishing and Submission
✔ Registration with CTSC
✔ Preparing IND package including Cover letter, IND, 1571, 1572 form and certification form 3674.
✔ Assembling and binding volumes
✔ Submission
https://www.creative-biolabs.com/immuno-oncology/ind-publishing-and-submission.htm
Pre-IND Meeting
For the drug development, pre-IND meeting is a critical tool to discuss the needs and challenges specific to the general product development, nonclinical testing, manufacturing information, protocol design or other regulatory questions defined in the Code of Federal Regulations (21 CFR 312.82). https://www.creative-biolabs.com/immuno-oncology/pre-ind-meeting.htm
Medical Writing and Translation
✔ Clinical overviews (eCTD Module 2.5) including literature review and references
✔ Clinical summaries (eCTD Module 2.7) including clinical pharmacology, efficacy, and safety
✔ Clinical study report preparation and review (eCTD Module 5)
✔ Clinical justification documents for EU, US and other emerging Regulatory markets
✔ Gap analysis for dossiers in clinical module
✔ Clinical and nonclinical document support, handling queries during HA meetings and responding to them
✔ Technical review dossiers
✔ Biowaiver support and justification document services
https://www.creative-biolabs.com/immuno-oncology/medical-writing-and-translation.htm
Regulatory Strategy Consulting
To gain approval for clinical testing after finalizing the pre-clinical testing of innovative new therapies, it is a key milestone for pharmaceutical companies to apply for approval of Investigational New Drug (IND) with FDA or other agencies.https://www.creative-biolabs.com/immuno-oncology/regulatory-strategy-consulting.htm
IND Regulatory Services
Navigating the drug development process from early stage discovery to clinical stage is complex and expensive. https://www.creative-biolabs.com/immuno-oncology/ind-regulatory-services.htm
cGMP Manufacturing
In the United States, the Current Good Manufacturing Practice (cGMP) is the Food and Drug Administration (FDA) 's formal regulations on the design, monitoring, control and maintenance of manufacturing processes and facilities.https://www.creative-biolabs.com/immuno-oncology/cgmp-manufacturing.htm
Antibody and Protein Toxicology
Pre-clinical toxicology is a study of the toxic effects of drugs in development based on statistical and quantitative analysis. https://www.creative-biolabs.com/immuno-oncology/antibody-and-protein-toxicology.htm
Antibody and Protein Pharmacology
Pharmacology is a key component concerned with the study of drug action in animal models which is essential and determinant to IND approval and ultimate NDA approval for a drug candidate. https://www.creative-biolabs.com/immuno-oncology/antibody-and-protein-pharmacology.htm
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Breast cancer in postmenopausal women with hormone receptor-positive (HR+) status is a common and complex condition that necessitates a multifaceted approach to management. HR+ breast cancer means that the cancer cells grow in response to hormones such as estrogen and progesterone. This subtype is prevalent among postmenopausal women and typically exhibits a more indolent course compared to other forms of breast cancer, which allows for a variety of treatment options.
Diagnosis and Staging
The diagnosis of HR+ breast cancer begins with clinical evaluation, imaging, and biopsy. Imaging modalities such as mammography, ultrasound, and MRI help in assessing the extent of the disease. Histopathological examination and immunohistochemical staining of the biopsy sample confirm the diagnosis and hormone receptor status by identifying the presence of estrogen receptors (ER) and progesterone receptors (PR) on the tumor cells.
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Phage display technology
- 1. Phage Display Technology in Creative Biolabs Creative Biolabs is a leading service provider of phage display library construction and screening. In a phage display library, a variety of peptides, small antibodies [e.g. scFv and Fab] or proteins are displayed on the surface of filamentous phage [M13, fd, and f1 strains] as fusion proteins with one of the coat proteins of the phage virions, while the genetic materials encoding the peptides/proteins are housed within the virions. Using a binding-based process called biopanning, a small number of phages that display proteins/peptides specifically binding to a target of interest can be rescued from a phage library that usually displays a repertoire of many billions of unique peptides/proteins. Finally, the peptides/proteins displayed by the selected phages can be identified by phage amplification and DNA sequencing. Phage Display Technology Phage Display Systems in Creative Biolabs M13 Phage Display pVIII-fusion display: major coat protein fusion, display up to 2700 copies of foreign protein pIII-fusion display: minor protein, display 1-5 copies of foreign protein Most popular option for phage display, has been applied in many different research areas. T4 Phage Display Larger genome DNA which enables larger insertions Dual display: two different molecules can be displayed separately on HOC and SOC Both N- and C-terminal insertion available T7 Phage Display Time saving: T7 phages have a shorter lifecycle than filamentous phages and lambda phages. Optimized biopanning: as T7 phages are resistant to extreme conditions, a variety of agents can be applied in screening procedure in contrast to alternative phages. Complementary to M13 phage, widely used for cDNA library. Library Construction Service in Creative Biolabs Phage Display Library Types Antibody library (immune, naïve, semi-synthetic, synthetic) Peptide library (linear, cyclic) Protein scaffold library cDNA Library Mutagenesis Strategies for Library Construction Trimer codon method Mutations are introduced at the codon level rather than at individual bases No codon bias, no frame shift, no stop codon Defined AA composition at each position Kunkel-like oligonucleotide-directed mutagenesis method Degenerate codon method Features No system-based out-of-frame mutations No limit on target length Flexible and scalable Large library capacity: from 108 to over 1011 Premade Antibody Libraries Available! During the past decades, we have integrated a comprehensive portfolio of premade libraries, including linear and cyclic peptide libraries, monobody libraries human scFv libraries, human Fab libraries, mouse scFv/Fab libraries, rabbit Fab libraries, chicken scFv libraries and single domain antibody libraries. Library Screening Service in Creative Biolabs Tailored Biopanning Strategies: Solid-phase screening Phage libraries are selected by flowing through a solid surface with the immobilized target. In-solution screening Isolating binders recognize naïve targets. The target-binder interaction is carried out in solution with subsequent capture by the appropriate method. Cell-based screening It is suitable to select peptides/antibodies for cell surface receptors, such as GPCR and ion channel-linked receptor. In Vivo screening Isolating novel peptides as the functional markers of new receptors or novel drug target candidates. Applications of Phage Display Technology Creative Biolabs offers high-quality phage display library construction and custom phage display library screening services for a broad range of project objectives including but not limited to: Function peptide discovery Therapeutic antibody discovery Monoclonal antibody discovery from a variety of species includes: human, monkey, llama, camel, shark, alligator, mouse, rat, hamster, guinea pig, rabbit, chicken, dog, bovine, goat, sheep, and ferret. Antibody humanization Antibody affinity maturation Contact Us Email: info@creative-biolabs.com Web: www.creative-biolabs.com Tel: 1-631-381-2994 Fax: 1-631-207-8356 Address: 45-1 Ramsey Road, Shirley, NY 11967, USA