A kind of nucleic acid molecule for preparing humanized antibody and its application
A nucleic acid molecule and carrier technology, which is applied to nucleic acid molecules and their application in the preparation of humanized antibodies to achieve high efficiency
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Embodiment 1
[0043] Transfer the modified human immunoglobulin gene into mice, and then immunize mice containing human immunoglobulin genes to obtain fully humanized antibodies. The specific steps are as follows:
[0044] 1. Optimization and transformation of human immunoglobulin genes
[0045] 1) Transformation of human immunoglobulin heavy chain gene
[0046] The human CD79 gene is transferred to the YAC or BAC vector containing human Ig through homologous recombination, and the gene cluster of the human immunoglobulin heavy chain gene is constructed as follows: figure 2 Shown (the position in the black box is the sequence of the imported human CD79 gene). Including the V region, D region and J region of the human immunoglobulin heavy chain gene, hIgHC μ, hIgHC δ, hIgHC γ3, hIgHC γ1, hIgHC α1, hLCR, hCD79. The internal structure of hCD79 is as follows Picture 1-1 As shown, its gene sequence is shown in SED ID NO.1; including the coding sequence of CD79α protein peptide and CD79β prot...
Embodiment 2
[0143] The transformed human immunoglobulin gene was transferred into a pig as a host animal (immunoglobulin gene knockout pig) to obtain the following results:
[0144] Detection of human IgG expression in pig blood: Two transgenic pigs, 205 and 206, which were positive in PCR detection, were selected to collect blood, separate serum, and detect the expression of human IgG in blood. as comparison. The antibodies identified by ELISA were: sigma(I1886) and sigma(A8792).
[0145] The result is as follows:
[0146] 4115: 0.5mg / ml
[0147] 4116: 0.1mg / ml
[0148] 4117: 0.2mg / ml
[0149] Healthy adults: 10.2mg / ml
[0150] Ordinary pig: <0.1ng / ml
[0151] The results show that the transferred gene can express human antibody heavy chain protein in pigs.
[0152] The modification scheme of the human immunoglobulin gene of the present invention is also applicable to other mammals, for example, pigs, rabbits, sheep, horses, etc. as host animals can also achieve the beneficial eff...
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