Preparation method of HRPII protein monoclonal antibody of plasmodium falciparum

A technology of Plasmodium falciparum and monoclonal antibody, which is applied in the fields of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of false positive test results, difficulty in preparing monoclonal antibodies, and difficulty in expression, etc. To achieve the effect of high specificity, enhanced stimulation, and improved expression level

CN101659975AActive Publication Date: 2010-03-03杭州新脉生物科技有限公司

0 Cites 14 Cited by

Patent Information

Authority / Receiving Office: CN · China
Current Assignee / Owner: 杭州新脉生物科技有限公司
Publication Date: 2010-03-03

Smart Images

Figure 1

Patent Text Reader

Abstract

The invention relates to a preparation method of HRPII protein monoclonal antibody of plasmodium falciparum. The preparation method comprises the following steps of: adopting HRPII protein of plasmodium falciparum as target antigen and respectively analyzing and selecting two dominant antigen epitopes of A and B; respectively repeating the two dominant antigen epitopes of A and B, then continuously connecting four glycine and forming recombinant protein C; adopting most securest code of escherichia coli and converting the amino acid sequence of the recombinant protein C into corresponding nucleotide sequence; carrying out chemical synthesis to the former step to obtain the nucleotide sequence, and respectively adding enzyme cutting sites BamHI and EcoRI at the upstream and downstream thereof; inserting nucleotide fragment obtained by the former step into expression carrier PET-28a(+), constructing recombinant protein C expression carrier and inducing to express the recombinant proteinC in the escherichia coli BL21 (DE3); carrying out ultrasonic bacteria breaking and low-temperature centrifugation, then taking supernatant of the solution, affining a chromatographic column by nickel-agarose, eluting and obtaining purified recombinant protein C; after immunizing Balb / c mouse with the recombinant protein C for a plurality of times, taking and fusing spleen cells with sp2 / 0 myelomacells, and obtaining six hybridoma cell lines by multiple rounds of screening; and purifying monoclonal antibody, respectively marking horse radish peroxidase and prorating matching and combination of optimum monoclonal antibody by ELISA orthogonal experiment.

Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The present invention relates to a method for tandem expression of dominant antigenic epitope of Plasmodium falciparum HRPII protein, preparation of monoclonal antibody to dominant antigenic epitope of Plasmodium falciparum HRPII protein and method for detection of Plasmodium falciparum by enzyme-linked immunosorbent assay The invention relates to a preparation method of HRPII protein monoclonal antibody, which belongs to the field of bioengineering technology manufacturing. Background technique

[0002] At present, due to the influence of various factors, no effective malaria vaccine has been put into practical use so far, so it is particularly important to realize rapid diagnosis and early treatment of malaria. Although the traditional thick and thin blood film smear microscopy method is specific, it is time-consuming and laborious, and requires skilled operators, so this method is not suitable for the rapid diagnosis of malaria. The sensitivity of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine

Login to View More