This document discusses protein fingerprinting (peptide mapping) as an analytical technique for protein identification. It involves cleaving a protein into peptides, separating the peptides, and analyzing them to generate a unique map or "fingerprint" for that protein. The technique was pioneered in the 1950s by Vernon Ingram to determine the difference between normal hemoglobin and sickle cell hemoglobin. He found that a single amino acid substitution was responsible for sickle cell disease. Peptide mapping is now commonly used in proteomics research to identify unknown proteins and compare similar proteins from different sources.

1. PROJECT REPORT ON
PROTIEN FINGERPRINTING(PEPTIDE MAPPING)
A Combined Dissertation Submitted in
Partial Fulfillment for The
AISSCE 2021-2022
GUIDED BY SUBMITTED BY:
DR.POLASHREE KHAUND ANKITA GOUR
PGT,BIOTECHNOLOGY CLASS-XII-B(SCIENCE)

2. ACKNOWLEDGEMENT
I would like to express my special thanks of gratitude to
Principal Shree Rajib Das Sir as for giving me this golden
opportunity to do this project on the topic "PROTIEN
FINGERPRINTING(PEPTIDE MAPPING)" which also
helped me in doing a lot of research I would like to
convey my sincere thanks to my teacher Dr. Polashree
Khaund (PGT Biotechnology )and Vrushali Sable(PGT
Biology) for her valuable suggestions and guidance in
completing this project work. I am indeed indebted to
her as she helped me lot in enhancing the knowledge on
the subject and overcoming the problems and
difficulties. Thanks to our lab attendant, my family and to
my dear friends who helped in finalizing this project
within the limited time frame. All of them contributed in
their own way to help me in making this project.

3. CERTIFICATE
This is to certify that this project entitled Protien
Fingerprinting(Peptide Mapping) presented in this report
work is carried out by "Ankita Gour", a student of class
XII-B, Science of Kendriya Vidyalaya Duliajan, under my
guidance.
This work has been prepared as a fulfillment for the
AISSCE 21-2022 and she seemed to have full dedication
while preparing this project.
SIGNATURE SIGNATURE
SUBJECT TEACHER EXTERNAL EXAMINER
PRINCIPAL

4. INDEX
SL.NO TITLE
1 INTRODUCTION
2 HISTORY
3 FOUR MAJOR STEPS
a)Isolation and purification of
the protien.
b) Selective Cleavage of the
peptide bonds.
c) Chromatographic separation
of the peptides.
d) Analysis and identification
of the peptide.
4 Diagram
5 Outcomes
6 Applications

5. INTRODUCTION
PROTEIN FINGERPRINTING / PEPTIDE MAPPING
It is an analytical technique for plain identification in which the
unknown protein of interest is first cleaved into smaller
peptides. whose absolute masses can be accurately measured
by many known techniques. In recent times, the masses of
these peptides is measured by mass spectrometer such as
MALDI-TOF or ESI-TOF. therefore, this technique is now
popularly referred to as Peptide Mass Fingerprinting (PMF).

6. HISTORY
This technique was invented by VM. Ingram in 1957 in the
Laboratory of Molecular Biology (LMB) at Cambridge, UK.
Ingram, John A. Hunt, and Antony 0. W. Stretton determined
that the change in the haemoglobin molecule in sickle cell
disease and trait was the substitution of the glutamic acid in
position 6 of the it-chain of the normal protein by valine.
Ingram used electrophoresis and chromatography to show that
the amino acid sequence of normal human and sickle cell
anaemia haemoglobins differed due to a single substituted
amino acid residue. Much of this work was done with the
support of Max Perutz and Francis Crick. Ingram won the
William Allan Award from the American Society of Human
Genetics in 1967. This was the first time a researcher
demonstrated that a single amino acid exchange in a protein
can cause a disease or disorder. As a result, Vernon Ingram is
sometimes referred to as The father of Molecular Medicine."

15. STEP 6
The chromatograms are dried and stained with a suitable
visualization reagent like Ninhydrin wherein peptide containing
regions appear as orange yellow spots.
STEP 7
The peptide map for HbA and HbS are compared and Ingram
found that one peptide was differently placed in the HbS map.
STEP 8
On examining this peptide and determining its amino acid
sequence, Ingram found that it had a valine substitution for
glutamic acid in the peptide.

17. OUTCOME
As a result the peptide spots were spread out in a characteristic
map or "fingerprint" The fingerprints of HbA and HbS showed
that all peptides occupied identical positions except for one.
called peptide no 4. which appeared in a new position in the
HbS fingerprint It must therefore have a different structure and
will represent the portion of the polypeptide chains where the
chemical difference between the two proteins lies The
structures of these two peptides. the HbA and HbS no. 4
peptides. are shown here.

18. APPLICATIONS
Peptide mapping became a useful technique to compare similar
proteins from different sources Slowly the information became
too vast and computers were used to store this data into
databases so that homology searches could be made
Mass spectrometry is a rapidly growing field of protein analysis,
which is proving useful in the identification of proteins
separated by 2-D gel electrophoresis. The most common mass
spectrometry protein identification technique is called peptide
mass fingerprinting.
The use of a peptide•mass fingerprint is fairly widespread in
proteomic research.