Journal of Shanghai Jiao Tong University (Medical Science) ›› 2023, Vol. 43 ›› Issue (6): 718-727.doi: 10.3969/j.issn.1674-8115.2023.06.008
• Basic research • Previous Articles
YIN Zi(), LIAN Chaoyang, GAO Bo, TIAN Ying, HAO Qian(), YEAP Lengsiew()
Received:
2023-01-13
Accepted:
2023-06-05
Online:
2023-06-28
Published:
2023-06-28
Contact:
HAO Qian,YEAP Lengsiew
E-mail:yinzi981030@sjtu.edu.cn;haoqian_2019@shsmu.edu.cn;yeaplengsiew@shsmu.edu.cn
Supported by:
CLC Number:
YIN Zi, LIAN Chaoyang, GAO Bo, TIAN Ying, HAO Qian, YEAP Lengsiew. Enhancement of the neutralization ability resulting from a single amino acid change in the light chain of a chimeric antibody against SARS-CoV-2[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023, 43(6): 718-727.
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URL: https://xuebao.shsmu.edu.cn/EN/10.3969/j.issn.1674-8115.2023.06.008
Primer | Sequence (5'→3') |
---|---|
1mFK_Ⅰ | RGTGCAGATTTTCAGCTTCCTGCT |
1mFK_Ⅱ | TGGACATGAGGGCYCCTGCTCAGT |
1mFK_Ⅲ | CTSTGGTTGTCTGGTGTTGAYGGA |
1mFK_Ⅳ | GTTGCTGCTGCTGTGGCTTACA |
1mFK_Ⅴ | GTATCTGGTACCTGTGG |
1mFK_Ⅵ | TGCCTGTTAGGCTGTTGGTGCT |
1mFK_Ⅶ | GCTCAGTTCCTTGGTCTCCTGTTGC |
1mFK_Ⅷ | TGGGTGCTGCTGCTCTGGGT |
1mFK_Ⅸ | CAGTTCCTGTTTCTGTTARTGCTCTGG |
1mFK_Ⅹ | TGCTCTGGTTATATGGTGCTGATGGG |
1mFK_Ⅺ | GCTGTTTTGTATACCTGGG |
1mFK_Ⅻ | CTCAGCTCCTGTTGCTGTGGC |
1mFK_ | CCTCATATTTTTGCTGCTATGGG |
1mFK_ⅩⅣ | TGCTTTTCTGGATTTCAGCCTCCAG |
1mFK_ⅩⅤ | TCAACTTCTGCTCTTCCTGCTGTTC |
1mFK_ⅩⅥ | CTAGCTCYTCTCCTCAGYCTTCTTCTCCTC |
1mFK_ⅩⅦ | GTGTMTGGTGCTBRTGGG |
1mFK_ⅩⅧ | STGYTGHTGYTCTGG |
1mFK_ⅩⅨ | GGGTATCTGGTACCTGTGG |
1mRK | ACTGAGGCACCTCCAGATGTT |
1mFL_Ⅰ | ACTTATACTCTCTCTCCTGGCTCTC |
1mFL_Ⅱ | CTCTTCTTCTTCTTTGTTCTTCATTGCT |
1mRL | GTACCATYTGCCTTCCAGKCCACT |
Tab 1 Primer sequences of the first-round nested PCR for constructing mouse-origin antibody light chain gene library
Primer | Sequence (5'→3') |
---|---|
1mFK_Ⅰ | RGTGCAGATTTTCAGCTTCCTGCT |
1mFK_Ⅱ | TGGACATGAGGGCYCCTGCTCAGT |
1mFK_Ⅲ | CTSTGGTTGTCTGGTGTTGAYGGA |
1mFK_Ⅳ | GTTGCTGCTGCTGTGGCTTACA |
1mFK_Ⅴ | GTATCTGGTACCTGTGG |
1mFK_Ⅵ | TGCCTGTTAGGCTGTTGGTGCT |
1mFK_Ⅶ | GCTCAGTTCCTTGGTCTCCTGTTGC |
1mFK_Ⅷ | TGGGTGCTGCTGCTCTGGGT |
1mFK_Ⅸ | CAGTTCCTGTTTCTGTTARTGCTCTGG |
1mFK_Ⅹ | TGCTCTGGTTATATGGTGCTGATGGG |
1mFK_Ⅺ | GCTGTTTTGTATACCTGGG |
1mFK_Ⅻ | CTCAGCTCCTGTTGCTGTGGC |
1mFK_ | CCTCATATTTTTGCTGCTATGGG |
1mFK_ⅩⅣ | TGCTTTTCTGGATTTCAGCCTCCAG |
1mFK_ⅩⅤ | TCAACTTCTGCTCTTCCTGCTGTTC |
1mFK_ⅩⅥ | CTAGCTCYTCTCCTCAGYCTTCTTCTCCTC |
1mFK_ⅩⅦ | GTGTMTGGTGCTBRTGGG |
1mFK_ⅩⅧ | STGYTGHTGYTCTGG |
1mFK_ⅩⅨ | GGGTATCTGGTACCTGTGG |
1mRK | ACTGAGGCACCTCCAGATGTT |
1mFL_Ⅰ | ACTTATACTCTCTCTCCTGGCTCTC |
1mFL_Ⅱ | CTCTTCTTCTTCTTTGTTCTTCATTGCT |
1mRL | GTACCATYTGCCTTCCAGKCCACT |
Primer | Sequence (5'→3') |
---|---|
2mFK_Ⅰ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGAAAWTGTGCTCACCCAGTC |
2mFK_Ⅱ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAAATTGTTCTCACCCAGTC |
2mFK_Ⅲ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCRACATTGTGCTGACCCAATC |
2mFK_Ⅳ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGAAACAACTGTGACCCAGTC |
2mFK_Ⅴ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATATTGTGATGACSCAGGC |
2mFK_Ⅵ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCRRTRTTGTGATGACCCARAC |
2mFK_Ⅶ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATATCCAGATGACACAGAC |
2mFK_Ⅷ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATTGTGATGACMCAGTC |
2mFK_Ⅸ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCCAGATGACHCAGTC |
2mFK_Ⅹ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCTTGCTGACTCAGTC |
2mFK_Ⅺ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCACACAGGCTGCACTCTCCAA |
2mFK_Ⅻ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCACTGGAGAAACAACACAGGC |
2mFK_ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATGTCCAGATAACCCAG |
2mFK_ⅩⅣ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCCTGATGACCCAATC |
2mFK_ⅩⅤ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATGTTGTGGTGACTCAAAC |
2mFK_ⅩⅥ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCAATATCCAGGTGATCCAG |
2mRK_Ⅰ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTCAGCTCCAGCTTGGTCCC |
2mRK_Ⅱ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTTATTTCCAGTCTGGTCCC |
2mRK_Ⅲ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTKATTTCCARCTTKGTSCC |
2mFL_Ⅰ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCTTACTCAGCCAAGCTCT |
2mFL_Ⅱ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAGGCTGTTGTGACTCAG |
2mFL_Ⅲ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAACYTGTGCTCACTCAG |
2mRL_Ⅰ | AGTGACCGTGGGGTTGGCCTTGGGCTGACCTAGGACAGTCAGTTTGGTTCC |
2mRL_Ⅱ | AGTGACCGTGGGGTTGGCCTTGGGCTGACCTAGGACAGTGACCTTGGTTCC |
Tab 2 Primer sequences of the second-round nested PCR for constructing mouse-origin antibody light chain gene library
Primer | Sequence (5'→3') |
---|---|
2mFK_Ⅰ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGAAAWTGTGCTCACCCAGTC |
2mFK_Ⅱ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAAATTGTTCTCACCCAGTC |
2mFK_Ⅲ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCRACATTGTGCTGACCCAATC |
2mFK_Ⅳ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGAAACAACTGTGACCCAGTC |
2mFK_Ⅴ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATATTGTGATGACSCAGGC |
2mFK_Ⅵ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCRRTRTTGTGATGACCCARAC |
2mFK_Ⅶ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATATCCAGATGACACAGAC |
2mFK_Ⅷ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATTGTGATGACMCAGTC |
2mFK_Ⅸ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCCAGATGACHCAGTC |
2mFK_Ⅹ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCTTGCTGACTCAGTC |
2mFK_Ⅺ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCACACAGGCTGCACTCTCCAA |
2mFK_Ⅻ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCACTGGAGAAACAACACAGGC |
2mFK_ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATGTCCAGATAACCCAG |
2mFK_ⅩⅣ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGACATCCTGATGACCCAATC |
2mFK_ⅩⅤ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCGATGTTGTGGTGACTCAAAC |
2mFK_ⅩⅥ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCAATATCCAGGTGATCCAG |
2mRK_Ⅰ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTCAGCTCCAGCTTGGTCCC |
2mRK_Ⅱ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTTATTTCCAGTCTGGTCCC |
2mRK_Ⅲ | GATGAAGACAGATGGCGCCGCCACAGTTCGTTTKATTTCCARCTTKGTSCC |
2mFL_Ⅰ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCTTACTCAGCCAAGCTCT |
2mFL_Ⅱ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAGGCTGTTGTGACTCAG |
2mFL_Ⅲ | CTGCTGCTCTGGTTCCCAGGTTCCAGATGCCAACYTGTGCTCACTCAG |
2mRL_Ⅰ | AGTGACCGTGGGGTTGGCCTTGGGCTGACCTAGGACAGTCAGTTTGGTTCC |
2mRL_Ⅱ | AGTGACCGTGGGGTTGGCCTTGGGCTGACCTAGGACAGTGACCTTGGTTCC |
Primer | Sequence (5 ́→3 ́) |
---|---|
Adapter forward primer | TTCCCAGGTTCCAGATGCCAGGCTGTTGTGACTCAG |
F98-deletion reverse primer | TGACCTTGGTTCCTCCACCGAACACCCAATGGTTGCTGTA |
T76-mutation reverse primer | CCTTGGTTCCTCCACCGAACACCCAGAAATGGTTGCTGTACCATAG |
Adapter reverse primer | GGTTGGCCTTGGGCTGACCTAGGACAGTGACCTTGG |
Tab 3 Primer sequences for plasmid construction with single amino acid mutation in mouse-origin antibody light chain
Primer | Sequence (5 ́→3 ́) |
---|---|
Adapter forward primer | TTCCCAGGTTCCAGATGCCAGGCTGTTGTGACTCAG |
F98-deletion reverse primer | TGACCTTGGTTCCTCCACCGAACACCCAATGGTTGCTGTA |
T76-mutation reverse primer | CCTTGGTTCCTCCACCGAACACCCAGAAATGGTTGCTGTACCATAG |
Adapter reverse primer | GGTTGGCCTTGGGCTGACCTAGGACAGTGACCTTGG |
Fig 6 Analysis of single amino acid site changes in the light chain of high-affinity human-mouse chimeric antibody and purification of antibodies with single amino acid changes
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