The development of strategy for obtaining single-chain recombinant antibodies against cell-surface biomarkers on the example of human CD34
| Autor: | Iu. S. Nikolaiev, O. B. Gorbatiuk, M. V. Tsapenko |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Phage display
QH301-705.5 Chemistry Cell CD34 Biopanning Single chain QH426-470 General Biochemistry Genetics and Molecular Biology Recombinant antibodies medicine.anatomical_structure Biochemistry Genetics medicine phage display Biology (General) biopanning single-chain recombinant antibodies (ScFv) |
| Zdroj: | Biopolymers and Cell, Vol 26, Iss 6, Pp 492-498 (2010) |
| ISSN: | 1993-6842 0233-7657 |
| DOI: | 10.7124/bc.000179 |
| Popis: | 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 |
| Databáze: | OpenAIRE |
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