Protein A adsorption medium
An adsorption medium and protein technology, which is applied in other chemical processes, chemical instruments and methods, etc., can solve the problem that protein A immunoadsorption column products cannot be finally sterilized
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[0014] 1. Preparation of recombinant protein A
[0015] Obtain the gene coding sequence of recombinant protein A by the following method:
[0016] 1. Directly synthesize the full sequence (SEQ ID NO.2);
[0017] 2. Synthesize the coding sequence (SEQ ID NO.3) of the repeating unit constituting the recombinant protein A. There is an AccI restriction site at both ends of the sequence. Since the recognition sequence of AccI is a non-palindromic sequence, it contains repeating units After the DNA fragment of the coding sequence is digested by the enzyme, it can be connected end-to-end to obtain a DNA fragment containing the coding sequence of several repeating units (Nilsson B, et al. A synthetic IgG-binding domain based on staphylococcal protein A. Protein Engineering, vol.1, 1987, p107-113), and then select the gene coding sequence of recombinant protein A with the complete sequence (SEQ ID NO.2).
[0018] While obtaining the recombinant protein A coding sequence, add suitable...
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[0024] Embodiment: the preparation of recombinant protein A
[0025] Directly synthesize the DNA fragment (SEQ ID NO.3) containing the repeating unit coding sequence of recombinant protein A. After digestion with NdeI and XhoI, the above DNA fragment is connected into the vector pUC18, which is well known in the art, and the vector is digested with AccI , the resulting small fragments are connected end to end, and then connected to the carrier fragments to obtain a recombinant plasmid containing several repeating unit coding sequences, and the recombinant protein A coding sequence (SEQ ID NO. 2) for the recombinant plasmid, the recombinant protein A coding sequence was transferred to the prokaryotic expression plasmid pET22b well known in the art by digestion with NdeI and XhoI to obtain the recombinant expression plasmid. The recombinant expression plasmid was transformed into Escherichia coli expression strain BL21(DE3), and the obtained recombinant expression strains were...
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