The document discusses recombinant antibody production. It begins by describing antibody structure and function, including the five main antibody isotypes found in vivo. It then details the process of natural antibody production by B cells, including B cell development, antibody gene rearrangement, and class switching. Finally, it introduces recombinant antibody production using hybridoma technology and lists the technical advantages of Creative Biolabs' hybridoma platform for generating recombinant antibodies.

1. www.creativebiolabs.net
Recombinant Antibody
Overview I

2. Contents
Recombinant Antibody Expression
Antibody Basics, Structure and Function
Antibody Production In Vivo
Recombinant Antibody Introduction
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3. An antibody is a large Y-shaped protein produced by B cells.
Recruited by the immune system to identify and neutralize foreign objects.
Antibody; Immunoglobulins Paratope of antibody
1.1 What is an Antibody?
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4. Antibody
Isotype
Type of Heavy
Chain
Sub-isotypes Function
IgA α IgA1, IgA2
Preventing pathogens invading into tissues
from epiderm
IgD δ N/A
Activating basophils and mast cells to
produce antimicrobial factors
IgE ε N/A
Interacting with allergens, facilitating
histamine releasing from mast cells and
basophils, preventing worms infection
IgG γ
IgG1, IgG2, IgG3,
IgG4
The major antibody fighting against
pathogens
IgM μ N/A
Provide early elimination of pathogens
before the generation of sufficient IgG
IgA
IgD IgGIgE
IgM
1.2 Antibody Isotypes in vivo
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5. 1.3 Antibody (IgG) Structure and Function
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• Constant Domains
• Functional
• Activities
• Variable Domains
• Antigen-binding
• Activity

6. 1.4 Antibody – Antigen Interaction
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Each antibody binds only to a specific antigen.
Antigen-antibody interactions are non covalent in nature.
The hinge region allows better binding
Four types of noncovalent forces

7. Contents
Recombinant Antibody Expression
Antibody Basics, Structure and Function
Antibody Production In Vivo
Recombinant Antibody Introduction
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8. 2.1 B lymphocytes Development
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Hematopoietic stem cell
Lymphoid cell
Pro-B cell
Pre-B cell
Immature B cell
Mature B cell
Partial heavy-chain gene
rearrangment
Complete heavy-chain gene rearrangment
Light-chain gene rearrangment
Change in RNA processing
CELL Ig EXPRESSED
None
None
None
μ Heavy chain
+surrogate light chain
mIgM
mIgM + mIgD
Bone marrow

9. 2.1 B lymphocytes Development
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Class switching
Memory B cells of various
isotypes
IgM
Differentiation
IgM-secreting plasma cells
Antigen stimulation
Plasma cells
secreting various
isotype
IgG IgA IgE
mIgM + mIgD
Peripheral lymphoid
organs
Mature B cell
Activated B cell

10. 2.2 Antibody Gene Rearrangement
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Primary transcript
RNA
Splicing
Light chain Heavy chain
Somatic recombination
Germline DNA
Somatic recombination
D-J rearranged DNA
joined
V-J or DJ joined
rearranged DNA
Transcription
DNARNAProtein
Polypeptide chain
mRNA
Translation

11. 2.3 Heavy Chain Locus and VDJ Rearrangement
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12. 2.4 Kappa Light Chain Rearrangement
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13. 2.5 Mechanism of Variable-Region DNA Rearrangements
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Recombination Signal Sequences (RSSs)
V(D)J recombinase = RAG1 and RAG2
Looping out of DNA
during rearrangement
12 base pairs (bp) – 1 turn of DNA
23 bp – 2 turns of DNA
12 can only join to 23 and vice versa
Between V, D, and J segments
Signal for recombination
2 kinds

14. 2.6 Antibody Production via Creative Biolabs’ Hybridoma Platform
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TECHNICAL ADVANTAGES
• Multiple-specie hybridoma generation;
• High immunization success rate: 90%;
• High screening capacity;
• High positive clone rate;
• Comprehensive down-stream process
development options.

15. Contact Us for Recombinant Antibody
Address: 45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-381-2994 Fax: 1-631-614-7828
Email: info@creative-biolabs.com
Web: www.creativebiolabs.net