Affinity Capture and Separation
From Molecular Interactions to Diagnostic Medical Applications
Norberto A. Guzman, Ph.D., Princeton Biochemicals Inc. Princeton, New Jersey, U.S.A.
Abstract
Proteomics in disease is highly complex due to the dynamic and diverse nature of proteins,
considered to have a greater chemical composition and structural function than any other biological
active compounds essential for life. Researchers have developed drafts of what is known as the
human proteome: the entire set of proteins expressed in the body, to help understand disease,
develop diagnostic tools, and accelerate precision medicine.
The human proteome is composed of subproteomes because each cell type has its own unique
proteome. Then there are proteoforms, which make up a proteome, and these proteoforms are the
protein variants - or forms - produced by a genome. Unlike the human genome, identifying and
studying an entire set of expressed proteins in the human body and organism poses a challenge to
scientists.
Proteomics techniques to study diseases focus on identifying, quantifying, and mapping protein
changes to discover biomarkers, understand mechanisms, and develop therapeutics. Due to the
complexity of the proteome, no single technique is adequate for a complete analysis of the
constituents. While existing methods provide valuable information, their limitations drive the
development of complementary, innovative methods to achieve greater breath of coverage,
dynamic range, or specificity of analysis.
Affinity capture techniques exploiting the specific binding between two molecules has been
employed for numerous purposes, from selective removal of interfering (over)abundant proteins
or enrichment of scarce biomarkers in complex biological samples to mapping the posttranslational
modifications (PTMs) and protein interactions with other proteins, nucleic acids or
biologically active small molecules.
In this seminar, I will discuss affinity capture separation techniques and focus on their unique
advantages for the selective enrichment of low-abundance proteins, as biomarkers of diseases,
employing antibodies, lectins and aptamers as affinity capture ligands immobilized to a surface
and coupled to capillary electrophoresis for separation of bound and eluted proteins and peptides
from complex biological samples (plasma, urine, saliva, sputum, tissues). On a final note, I will
outline emerging methods that offer significant promise to proteomics research.
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