Composition containing group B meningococcal fHBP antigen as well as preparation method and application thereof
A meningococcal and composition technology, applied in the field of biomedicine, can solve problems such as insufficient immunogenicity, and achieve the effects of facilitating expression and purification, widening antigen spectrum, and improving immune effect
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[0029] According to one aspect of the present invention, the present invention also relates to a method for preparing a composition comprising group B meningococcal fHBP antigen as described above, comprising the following steps:
[0030] Obtaining recombinant plasmids, which respectively contain coding genes capable of expressing V1, V2 and V3;
[0031] Transforming the recombinant plasmids into host cells respectively, identifying, screening and identifying correct positive strains, and transforming the correctly identified positive strains into expression cells; culturing;
[0032] V1, V2 and V3 are isolated and enriched from the expressing cells and mixed with an optional adjuvant.
[0033] In some embodiments, the encoding genes capable of expressing V1, V2 and V3 are:
[0034] a) sequentially as shown in SEQ ID NO: 4-6; or
[0035] b). A sequence that encodes the same protein as the nucleotide sequence of a), but differs from the nucleotide sequence of a) due to codon ...
Embodiment
[0054] 1. Construction of recombinant plasmids
[0055] Three fHBP genes with P4 signal peptide were artificially synthesized by means of genetic engineering.
[0056] B01(V1)-fHBP nucleotide sequence (the italic bold part is the P4 signal peptide nucleotide sequence):
[0057]
[0058] The expressed protein sequence is as follows (the bold part in italics is the amino acid sequence of the P4 signal peptide, and the rest corresponds to SEQ ID NO: 1):
[0059]
[0060] A19(V2)-fHBP nucleotide sequence (the italic bold part is the P4 signal peptide nucleotide sequence):
[0061]
[0062] The expressed protein sequence is as follows (the bold italic part is the amino acid sequence of the P4 signal peptide, and the rest corresponds to SEQ ID NO:2):
[0063]
[0064] A05(V3)-fHBP nucleotide sequence (the italic bold part is the P4 signal peptide nucleotide sequence)
[0065]
[0066] The expressed protein sequence is as follows (the bold part in italics is the ami...
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