Copyright
©The Author(s) 2004.
World J Gastroenterol. Sep 15, 2004; 10(18): 2619-2623
Published online Sep 15, 2004. doi: 10.3748/wjg.v10.i18.2619
Published online Sep 15, 2004. doi: 10.3748/wjg.v10.i18.2619
Table 1 Oligonucleotide primers used for PCR of human immunoglobulin genes
| A. Primary PCRs | |
| Human VH back primers (sense) | |
| HuVHlaBACK | 5’-CAGGTGCAGCTGGTGCAGTCTGG-3’ |
| HuVH2aBACK | 5’-CAGGTCAACTTAAGGGAGTCTGG-3’ |
| HuVH3aBACK | 5’-GAGGTGCAGCTGGTGGAGTCTGG-3’ |
| HuVH4aBACk | 5’-CAGGTGCAGCTGCAGGAGTCGGG-3’ |
| HuVH5aBACK | 5’-GAGGTGCAGCTGTTGCAGTCTGC-3’ |
| HuVH6aBACK | 5’-CAGGTACAGCTGCAGCAGTCAGG-3’ |
| JH forward primers (anti-sense) | |
| HuJHl-2FOR | 5’-TGAGGAGACGGTGACCAGGGTGCC-3’ |
| HuJH3FOR | 5’-TGAAGAGACGGTGACCATTGTCCC-3’ |
| HuJH4-5FOR | 5’-TGAGGAGACGGTGACCAGGGTTCC-3’ |
| HuJH6FOR | 5’-TGAGGAGACGGTGACCGTGGTCCC-3’ |
| Human V kappa back primers (sense) | |
| HuV κ laBack | 5’-GACATCCAGATGACCCAGTCTCC-3’ |
| HuV κ 2aBack | 5’-GATGTTGTGATGACTCAGTCTCC-3’ |
| HuV κ 3aBack | 5’-GAAATTGTGTTGACGCAGTCTCC-3’ |
| HuV κ 4aBack | 5’-GACATCGTGATGACCCAGTCTCC-3’ |
| HuV κ 5aBack | 5’-GAAACGACACTCACGCAGTCTCC-3’ |
| HuV κ 6aBack | 5’-GAAATTGTGCTGACTCAGTCTCC-3’ |
| Human J kappa forward primer (anti-sense) | |
| HuJ κ 1FOR | 5’-ACGTTTGATTTCCACCTTGGTCCC-3’ |
| HuJ κ 2FOR | 5’-ACGTTTGATCTCCAGCTTGGTCCC-3’ |
| HuJ κ 3FOR | 5’-ACGTTTGATATCCACTTTGGTCCC-3’ |
| HuJ κ 4FOR | 5’-ACGTTTGATCTCCACCTTGGTCCC-3’ |
| HuJ κ 5FOR | 5’-ACGTTTAATCTCCAGTCGTGTCCC-3’ |
| Human V lambda back primers (sense) | |
| HuV λ 1BACK | 5’-CAGTCTGTGTTGACGCAGCCGCC-3’ |
| HuV λ 2BACK | 5’-CAGTCTGCCCTGACTCAGCCTGC-3’ |
| HuV λ 3aBACK | 5’-TCCTATGTGCTGACTCAGCCACC-3’ |
| HuV λ 3bBACK | 5’-TCTTCTGAGCTGACTCAGGACCC-3’ |
| HuV λ 4BACK | 5’-CACGTTATACTGACTCAACCGCC-3’ |
| HuV λ 5BACK | 5’-CAGGCTGTGCTCACTCAGCCGTC-3’ |
| HuV λ 6BACK | 5’-AATTTTATGCTGACTCAGCCCCA-3’ |
| Human J lambda forward primers (anti-sense) | |
| HuJ λ 1FOR | 5’-ACCTAGGACGGTGACCTTGGTCCC-3’ |
| HuJ λ 2-3FOR | 5’-ACCTAGGACGGTCAGCTTGGTCCC-3’ |
| HuJ λ 4-5FOR | 5’-ACCTAAAACGGTGAGCTGGGTCCC-3’ |
| B. PCR assembly | |
| Hu JH-Linker primers | |
| HuJH1-2Linker | 5’-AGAGCCACCTCCGCCTGAACCGCCTCCACCTGAGGAGACGGTGACCAGGGTGCC-3’ |
| HuJH3Linker | 5’-AGAGCCACCTCCGCCTGAACCGCCTCCACCTGAAGAGACGGTGACCATTGTCCC-3’ |
| HuJH4-5Linker | 5’-AGAGCCACCTCCGCCTGAACCGCCTCCACCTGAGGAGACGGTGACCAGGGTTCC-3’ |
| HuJH6Linker | 5’-AGAGCCACCTCCGCCTGAACCGCCTCCACCTGAGGAGACGGTGACCGTGGTCCC-3’ |
| Linker-Hu V κ primers | |
| Linker-HuV κ 2-3-6 BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAWRTTGTGHTGACKCAGTCTCC-3’ |
| Linker-HuV κ 1-4 BACK | 5’-gTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCSWGATGACCCAGTCTCC-3’ |
| Linker-HuV κ 5 BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAAACGACACTCACGCAGTCTCC-3’ |
| Linker-Hu V λ primers | |
| Linker-HuV λ 1-2BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGCAGTCTGYSYTGACKCAGCCKSC-3’ |
| Linker-HuV λ 3BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGTCYTMTGWGCTGACTCAGSMMCC-3’ |
| Linker-HuV λ 4-5BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGCASGYTRTRCTSACTCARCCGYC-3’ |
| Linker-HuV λ 6BACK | 5’-GTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGAATTTTATGCTGACTCAGCCCCA-3’ |
| C. Reamplification with primers containing restriction sites | |
| Human VH back (Sfi) primers (sense) | |
| HuVH1aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTGCAGCTGGTGCAGTCTGG-3’ |
| HuVH2aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTCAACTTAAGGGAGTCTGG-3’ |
| HuVH3aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCGAGGTGCAGCTGGTGGAGTCTGG-3’ |
| HuVH4aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTGCAGCTGCAGGAGTCGGG-3’ |
| HuVH5aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTGCAGCTGTTGCAGTCTGC-3’ |
| HuVH6aBACKSfi | 5’-GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCCCAGGTACAGCTGCAGCAGTCAGG-3’ |
| Human J kappa forward (Not) primers (anti-sense) | |
| HuJ κ 1FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTGATTTCCACCTTGGTCCC-3’ |
| HuJ κ 2FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTGATCTCCAGCTTGGTCCC-3’ |
| HuJ κ 3FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTGATATCCACTTTGGTCCC-3’ |
| HuJ κ 4FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTGATCTCCACCTTGGTCCC-3’ |
| HuJ κ 5FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACGTTTAATCTCCAGTCGTGTCCC-3’ |
| Human J lambda forward (Not) primers (anti-sense) | |
| HuJ λ 1FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACCTAGGACGGTGACCTTGGTCCC-3’ |
| HuJ λ 2-3FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACCTAGGACGGTCAGCTTGGTCCC-3’ |
| HuJ λ 4-5FORNot | 5’-GAGTCATTCTCGACTTGCGGCCGCACCTAAAACGGTGAGCTGGGTCCC-3’ |
Table 2 Immunore activity of soluble scFv determined by ELISA
- Citation: Xu MY, Xu XH, Chen GZ, Deng XL, Li J, Yu XJ, Chen MZ. Production of a human single-chain variable fragment antibody against esophageal carcinoma. World J Gastroenterol 2004; 10(18): 2619-2623
- URL: https://www.wjgnet.com/1007-9327/full/v10/i18/2619.htm
- DOI: https://dx.doi.org/10.3748/wjg.v10.i18.2619