Biopolymers and cell. 2010; 26 (6): 492 - 498
The development of strategy for obtaining single-chain recombinant antibodies against cell-surface biomarkers on the example of human CD34
Iu. S. Nikolaiev, O. B. Gorbatiuk1, M. V.Tsapenko
State Organization of Genetic and Regenerative Medicine NAMS of Ukraine, 67, Vyshgorodska Str., Kyiv, Ukraine, 04114
1Taras Shevchenko National University of Kyiv 64, Volodymyrska Str., Kyiv, Ukraine, 01033
Antibodies against cell-surface proteins play an important role in cell detection, separation and determination of differentiation stage. The globular structure of extracellular region of cell-surface antigens is frequently characterized by heavily glycosilation and/or is stabilized by disulphide bonds. In this case the obtaining of antibodies against such proteins is a substantive problem. Aim. The development of strategy for obtaining recombinant antibodies against cell-surface biomarkers. Methods. The research strategy is based on the construction of cDNA library of VH and VL genes of animals, immunized with recombinant antigen, and subsequent cell-based biopanning for the library enrichment with desired phage clones. High-throughput automated systems were used for the detection and isolation of antigen-specific clones. Results. We have obtained a panel of five antibodies that recognize an antigen on CD34+ cell surface using the methods of immunocytochemistry and flow cytometry. Conclusions. The proposed strategy may be used for obtaining antibodies against cell-surface antigens.
Keywords: CD34, phage display, single-chain recombinant antibodies (ScFv), biopanning.