Protein Analyzing Approaches and Its Application to Immunization

Introduction to Protein

Proteins are large biomolecules, or polymers, which consist of one or more long
chains consisting of amino acid residues. The amino acid molecules are arranged in a
linear arrangement, and the carboxyl groups and amino groups of the adjacent amino
acid residues are linked by peptide bonds. Like other biological macromolecules (such
as polysaccharides and nucleic acids), protein is a necessary component of the
organism on the earth getting involved in every process of metabolism. The amino
acid sequence of proteins is encoded by corresponding genes, which means that
protein is external expression of DNA (deoxyribonucleic acid).

Protein Analyzing Approaches

1. Amino Acid Analysis

If we need to further apply the knowledge of the quantities of protein or peptide,

quantitative amino acid analysis (qAAA) will be a proper assay, which can not only
measure the specific protein quantities, but provide detailed relevant information,
There are 4 steps of qAAA procedure: hydrolysis, separation, detection and
quantification.

2. Mass Spectrometry

(1) de-novo protein sequence analysis

Without the database, de-novo protein sequence analysis directly translates the amino
sequences from tandem mass spectra. We ought to attach great importance to this kind
of way because de-novo protein sequence analysis plays an essential role when
detecting unknown protein sequence.
(2) database searching

Protein database is a database that includes protein information. Normally, when

dealing with a bunch of protein, searching the information in an authoritative database
should occur to us at the first time, as it is the most convenient and efficient way.
There are many protein databases in common use, among which Uniprot is considered
to be the most extensive protein database with the most comprehensive annotation.
For instance, shotgun proteomics is one of the most widely-used approaches to
confirm the sequence of the protein.

Application of Protein Analysis to Immunization

Introduction to Immunization

Immunization is a physiological function of the human body. The human body relies
on this function to identify if the substance inside the body belong to itself, thereby
destroying and repelling the antigenic substances (such as germs, viruses, etc.) or the
damaged cells and tumor cells produced by the human body to maintain people’s
health.

1. Host Cell Protein Analysis

In mankind history, human beings fight against virus for millenniums. About diseases
caused by viruses, there were records of smallpox in India and China during two to
three B. C. Viruses-caused diseases is detrimental to human’s health. AIDS, caused by
HIVs, a kind of RNA virus, is still an incurable disease for people. In the midst of the
rapid development of modern science and technologies, study on viruses is
indispensable for human, among which Host Cell Protein Analysis is one of the most
critical parts. Cells that are invaded by a specific virus are called host cells and Host
Cell Proteins (HCPs) are a heterogeneous, complex group of proteins that are derived
from the host cell. Identification and quantification of host cell proteins is of great
importance in the development of biopharmaceutical products to guarantee the purity
of products and patient safety.

2. De Novo Antibody Sequencing

Antibody is a large Y - shaped protein that is secreted by plasma cells (effect B cells)
and is used by the immune system to identify and neutralize foreign substances, such
as bacteria, viruses, and so on. Antibodies can recognize a unique feature of a specific
foreign object, which is called antigen. The unique biological characteristic of
antibodies makes them play an important role in the diagnosis, immunization and
basic research of diseases. Early in the late nineteenth Century, specific antigens were
used to immunize animals to prepare corresponding antisera. As we all know, most of
the antibodies consist of proteins. Therefore, De Novo Antibody Sequencing is a kind
of measure of de-novo protein sequence analysis.