Preparation method of latex microsphere cross-linking specific monoclonal or polyclonal antibody for detecting soluble abnormally glycosylated modified CD58 molecule
A polyclonal antibody, glycosylation technology, applied in the field of medical testing, can solve the problems of long detection time and low sensitivity
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[0024] In order to make the technical means, creative features, goals and effects achieved by the present invention easy to understand, the present invention is further described using examples.
[0025] 1. Preparation method of abnormally glycosylated CD58 molecule monoclonal IgG antibody
[0026] 1.1 Animal immunity
[0027] The total RNA of 1х106 colorectal cancer LoVo cells was isolated and purified with RNAeasy kit, the total RNA was reverse transcribed into cDNA by Oligo dT15 primer, the human CD58 gene sequence (NM_001779.2) was amplified by polymerase chain reaction (PCR), and analyzed by EcoRI After digestion with / BamH1, it was cloned into the pLXSN retroviral vector, verified by DNA sequencing, a CD58 expression retroviral vector system was constructed, transfected into PT67 cells to produce recombinant retroviruses, and infected with colorectal cancer SW620 cells at a titer of 10:1. Positive clones were screened for 7-14 days with 500-700 μg / mL of G418 drug pressu...
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