Monocytic leukemia associated antigen MLAA-34 resistant fully human monoclonal single-chain antibody ScFv

A mononuclear cell and single-chain antibody technology, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, immunoglobulin, etc., can solve the problem of non-panning And identified problems such as fully human monoclonal antibody, to achieve the effect of small molecular weight, easy application, and strong penetrating power

Active Publication Date: 2016-07-27
THE SECOND AFFILIATED HOSPITAL OF XIAN JIAOTONG UNIV
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AI Technical Summary

Benefits of technology

This patented technology provides an improved way to identify or isolate certain types of cancer cells called melanoma (melanosis). These can be found throughout our body's bloodstream but they are often very difficultly detected due to their similarity from other normal tissues such as skin and lymph nodes. To solve this issue, we developed two different ways: one uses full length monomers derived form fusion proteins containing both mouse A3a and B7-1 polypeptides; another involves creating artificial mAbs made up only of these parts instead of whole ones through DNA manipulation techniques like cloning them into bacteria.

Problems solved by technology

This patented describes various methods aim at identifying certain types of cancer called metastasis-positive breast cancer (MBSC)/myxoma/non-humans sarcoma (MMHSPC)-chromoonsilvate carboxyl transfer ribon translation enzyme complex(TRAP1), resulting from fusion between two complement regulatory Bcl-1 family members. These technologies provide tools for studying how these targets interact with each other during inflammatory responses caused by activator response mediators like histones and cytokines released through stimulation of Toll Like Receptor-4-5 adaptive immunity.

Method used

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  • Monocytic leukemia associated antigen MLAA-34 resistant fully human monoclonal single-chain antibody ScFv
  • Monocytic leukemia associated antigen MLAA-34 resistant fully human monoclonal single-chain antibody ScFv
  • Monocytic leukemia associated antigen MLAA-34 resistant fully human monoclonal single-chain antibody ScFv

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Embodiment 1

[0059] 1.1.1 Construction of prokaryotic expression vector

[0060] The total RNA in U937 cells was extracted, reverse-transcribed into cDNA, using cDNA as a template strand, primers were designed, and the coding region of MLAA-34 was amplified by PCR. The target carrier is digested, and the digested products are recovered by electrophoresis and exchanged, and the products are transformed into bacterial competent cells. Colony PCR identification was performed on the grown clones first, and then sequencing and comparative analysis were performed on the positive clones identified by PCR. The correct comparison was the successful construction of the target plasmid ( Figure 3-1 ). figure 1 Flowchart for the construction of the prokaryotic expression vector for the target gene, figure 2 A map of the expression vector.

[0061] 1) Enzyme digestion of the vector.

[0062] Enzyme digestion reaction temperature: 37°C, enzyme digestion reaction time: 2h,

[0063] Table 1 enzyme dige

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Abstract

The invention provides a monocytic leukemia associated antigen MLAA-34 resistant fully human monoclonal single-chain antibody ScFv. The monoclonal single-chain antibody ScFv of special anti-MLAA-34 is obtained by screening and identifying in a phage antibody library; MLAA-34 protein is firstly expressed and purified, and antigen protein is provided for monoclonal antibody preparation. A method beneficial to genetic engineering is used for screening and identifying anti-MLAA-34 fully human monoclonal antibody. The fully human monoclonal single-chain antibody ScFv for specially combining and efficiently expressing tumor cell strains of MLAA-34 is obtained. The fully human monoclonal single-chain antibody ScFv can be used for solving the immunogenicity problem of a mouse original antibody, and the single-chain antibody ScFv is formed by connecting a heavy chain and a light chain of an antibody variable region, is small in molecular weight, is high in penetrating power and is more beneficial to application.

Description

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Claims

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Application Information

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Owner THE SECOND AFFILIATED HOSPITAL OF XIAN JIAOTONG UNIV
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