Carcino-embryonic antigen monoclonal antibody, chip containing same and application
A carcinoembryonic antigen and antibody chip technology is applied in the field of bioengineering to achieve accurate qualitative detection, convenient operation and strong specificity
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Embodiment 1
[0044] Embodiment 1: Preparation of the monoclonal antibody of the present invention
[0045] The preparation method of antibody comprises the following process steps:
[0046] (1) Cell fusion: Sp2 / 0 myeloma cells with high activity were mixed with splenocyte suspensions sensitized with self-designed and synthesized carcinoembryonic antigen polypeptide (sequence TQQAT PGPAYSGREI) at a ratio of 1:10-100, and added Polyethylene glycol (produced by Sigma, USA) makes the cells fused with each other, and the culture medium is added dropwise to the mixed cell suspension of the two cells, and the cells are cultured with HAT selective medium (produced by Haiclone, USA);
[0047] (2) Screen hybridoma cells: when the cells to be fused are cultured to the 5th to 10th day, absorb the culture supernatant of the clonal cell clusters in the wells of the 96-well culture plate, and use the enzyme-linked immunosorbent assay method to detect the antibody content, and limit dilution Three times ...
Embodiment 2
[0068] Embodiment 2: the preparation method of antibody chip
[0069] Increase following processing steps on the basis of embodiment 1:
[0070] Step (5) Label complement C3 antibody with CY3 fluorescein.
[0071] Step (6) Spot 96 spots on one chip, two spots for each monoclonal antibody, and the content of each spot is 0.01-1ng / ml.
Embodiment 3
[0072] Embodiment 3: Detection of various indicators of the monoclonal antibody described in the present invention
[0073] The monoclonal antibody secreted by the hybridoma cell line screened in Example 1 was subjected to sequence analysis, and its heavy chain variable region was as follows:
[0074] VKLQESGPGL VAPSQSLSMS CTVSGFSLSS YGVHWVRQPPGKGLEWLGVI WAGGTTNYNS ALMSRLSISK DNSKSQVLLKMNSLQTDDTA MYYCATTTMI TLMDYWGQGT TVTVSSA (shown in SEQ ID No. 1)
[0075] Its light chain variable region is as follows:
[0076] DVQLNQAKSS LSASLGDRVT ISCRASQDIS NYLNWYQQKP DGTVKLLIYY TSRLHSGVPP RFSGSGSGTD YSLTISNLEQ DIATYFCQQGNTVPWTFGG GTKLEIS (shown in SEQ ID No. 2)
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