Anti-human st2 antibody and use thereof
An antibody and sequence technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, applications, etc., can solve the problems of no transmembrane domain and unclear functions
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0157] Example 1: Construction of phage display antibody library
[0158] For specific experimental procedures, please refer to Example 1 of Chinese Patent Application No. 201610609651.X and Example 1 of Chinese Patent Application No. 201510097117.0. The entire contents of the above two patent applications are incorporated herein by reference.
Embodiment 2
[0159] Embodiment 2: Preparation of recombinant protein
[0160] A variety of different recombinant proteins are used in the preparation of anti-ST2 monoclonal antibodies, including human ST2 extracellular region (hST2, SEQ ID NO:1), mouse ST2 extracellular region (mST2, SEQ ID NO:2) , macaque ST2 extracellular region (mmST2, SEQ ID NO:3) and human IL-33 extracellular region (hIL-33, SEQ ID NO:4). These proteins have a large number of post-translational modifications (such as glycosylation or disulfide bonds, etc.), so the use of mammalian cell expression systems will be more conducive to maintaining the structure and function of recombinant proteins. In addition, a His tag (His, SEQ ID NO:5) or the Fc fragment of human IgG1 (hFc, SEQ ID NO:6) or the Fc fragment of mouse IgG2a (mFc, SEQ ID NO:6) was added to the C-terminus of these recombinant proteins. NO:7), which will be more conducive to the purification of recombinant proteins and the identification of monoclonal antibod...
Embodiment 3
[0163] Example 3: Screening of anti-human ST2 monoclonal antibodies using phage display antibody library technology
[0164] 3.1 Screening of anti-human ST2 monoclonal antibody
[0165] Using the recombinant human ST2-his prepared in Example 2 as an antigen, a solid-phase screening strategy (experimental protocol refers to phage display: general experimental guidelines / (US) Clarkson (Clackson, T.), (US) Loman (Lowman, Edited by H.B.; Translated by Ma Lan et al. Chemical Industry Press, 2008.5) Screened the phage library displaying the human single-chain antibody library prepared in Example 1, and finally obtained a fully human single-chain antibody S1A4 that specifically binds to human ST2.
[0166] 3.2 Preparation of fully human antibody
[0167] Using conventional molecular biology methods, the nucleic acid molecules encoding the heavy chain variable region of S1A4 (S1A4 VH, SEQ ID NO:18) and the light chain variable region (S1A4 VK, SEQ ID NO:19) were respectively cloned i...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


