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Date:6/16/2010

antibodies. These proteinsproduced by the B cells of the body's immune systemare able to selectively bind with disease-associated antigens. Various techniques permit researchers to probe blood serum, pulling out the antigens that bind to the antibodies and observing them with the aid of fluorescence or other methods.

Traditional means of producing antibodies for research are labor-intensive and cumbersome, requiring injection of antigens into animals, which then act as production sites for the antibodies of interest. A new method however allows antibodies to be engineered synthetically. In this scenario, the portion of the antibody responsible for binding with an antigen, known as the variable region, is built from amino acids. These single chain variable fragments or scFvs can be inserted into bacteria, which act as vectors for the antibody fragments, much as a mouse or other animal would carry a traditional antibody.

scFvs encoded in bacteria have been assembled into vast libraries, capable of probing the full complexity of proteins found in blood. When an scFv detects something unusual in the blood that may suggest an abnormality, it binds to it, just as in a normal immune response. Ramirez used a library of scFvs to find which antibody fragments selectively attached to proteins in blood carrying ovarian cancer, eliminating the antibodies that bound with proteins in normal blood. The method yielded 19 distinct scFvs that appeared to display specific affinity for proteins exclusively found in ovarian cancerous blood serum.

Identification of the protein antigens bound to these scFvs however, proved far trickier. After several years of frustrating attempts to recognize these protein culprits, Ramirez teamed up with LaBaer, applying a new method that would allow them to screen the cancer-associated scFvs against thousands of proteins of known sequence.

The techniqueknown as NAPPA (for nucleic acid programmable protein array),
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Contact: Kimberly Ovitt
Kimberly.Ovitt@asu.edu
Arizona State University
Source:Eurekalert  

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