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Expression of recombinant anti‐colorectal cancer large single‐chain monoclonal antibody in insect cells
Author(s) -
Shao Yingxue,
Lee JeongHwan,
Choo YoungKug,
Ko Kisung
Publication year - 2012
Publication title -
entomological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.421
H-Index - 20
eISSN - 1748-5967
pISSN - 1738-2297
DOI - 10.1111/j.1748-5967.2012.00479.x
Subject(s) - recombinant dna , monoclonal antibody , biology , microbiology and biotechnology , colorectal cancer , sf9 , cancer cell , immunoglobulin light chain , western blot , antigen , antibody , virology , cancer research , cancer , gene , immunology , biochemistry , spodoptera , genetics
Colorectal cancer is the third most commonly diagnosed cancer in the world. Monoclonal antibody (m A b) CO 17‐1 A recognizes the tumor‐associated antigen GA 733‐2, a cell surface glycoprotein highly expressed in colorectal carcinoma cells which is applicable for preventing and curing colorectal cancer. In this study, we tried to produce a new recombinant anti‐colorectal cancer large single chain (lsc) m A b based on m A b CO 17‐1 A in the baculovirus‐insect cell protein expression system. Two kinds of recombinant lsc mAbs were generated where variable light chain ( VL ) and heavy chain ( HC ) of m A b CO 17‐1 A were fused together by an interchain linker. The only difference between the two m A bs is based on fusion of an ER retention signal ( KDEL ) at its C ‐terminus of HC . Polymerase chain reaction analysis verified the presence of both recombinant genes in the bacmid for generating viral expression vectors in insect cells. Western blot confirmed the expression of lsc mAbs in baculovirus‐infected insect cells. Cell enzyme linked immunosorbent assay ( ELISA ) showed that the m A bs from cell lysates bound to SW 480 and SW 620 human colorectal cancer cells. These results indicate that the baculovirus insect expression system can produce anti‐colorectal lsc m A b recognizing human colorectal cancer cells.