Human LP (legionella pneumophila) surface protein monoclonal antibody and application
A Legionella pneumophila, monoclonal antibody technology, applied in the field of immunology, can solve the problem that the role of outer membrane lipoprotein is not clear enough, and achieve good specificity and stability
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Embodiment 1
[0042] Example 1 Preparation of recombinant human Legionella pneumophila surface protein PAL and its polyclonal antibody
[0043] (1) Cloning and expression of human Legionella pneumophila PAL gene
[0044] Bioinformatic analysis of the human Legionella pneumophila surface protein PAL (the accession number in the NCBI protein database is YP_096059) gene, combined with GC content, codon bias, mRNA secondary structure, RNA instability motif, mRNA freedom Considering performance stability, etc., the DNA coding sequence was optimized, and at the same time, the whole gene sequence was chemically synthesized after introducing the restriction site NdeI at the 5' end, the termination signal TAA and the restriction site EcoRI at the 3' end (the whole sequence synthesis was handed over to Kim Completed by Sirui Biotechnology Co., Ltd., the artificially synthesized gene fragment was connected to the vector pUC57 at the time of delivery), denoted as PAL'. The full sequence of the gene an...
Embodiment 2
[0060] Example 2 Preparation of Human Legionella Pneumophila Surface Protein Monoclonal Antibody Lp-1#
[0061] 1) Immunization of animals
[0062]Using the recombinant human Legionella pneumophila surface protein PAL prepared in Example 1 as an antigen, 5 8-week-old BALB / c mice were immunized, and 2 were not used as immunized mice as negative controls. After the initial immunization antigen was fully emulsified with an equal amount of Freund's complete adjuvant, the mice were subcutaneously injected into the back of the immunized mice at a rate of 100 μg / mouse. After that, the second immunization was performed by intraperitoneal injection with the same dose of antigen fully emulsified with Freund's incomplete adjuvant at an interval of three weeks, and the third time was injected intraperitoneally with the same dose of antigen fully emulsified with Freund's incomplete adjuvant at an interval of 2 weeks. immunity. Blood was collected from the tail vein 15 days after the thir...
Embodiment 3
[0108] Example 3 Human Legionella pneumophila immunochromatographic detection test strip
[0109] Human Legionella pneumophila immunochromatographic detection test strip (such as figure 1 shown) is composed of sample pad 1, binding pad 2, detection layer 3, detection line 4, quality control line 5, water-absorbing pad 6, and rigid polyvinyl chloride backing plate 7;
[0110] The nitrocellulose membrane detection layer 3 is pasted on the rigid polyvinyl chloride backing plate 7, and the bonding pad 2 and the sample pad 1 are pasted on the left side of the nitrocellulose membrane detection layer 3 from bottom to top in sequence, and the water-absorbing pad 6 is arranged on the The position on the right side above the nitrocellulose membrane detection layer 3. The conjugation pad 2 is made of glass cellulose membrane, and colloidal gold-labeled monoclonal antibody Lp-1# (secreted by hybridoma cell line Lp-1#, preservation number CCTCC NO: C2017213) is sprayed on the conjugation ...
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