Antigen-binding molecule capable of binding to plurality of antigen molecules repeatedly
An antigen-binding molecule, binding domain technology, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, antibody, combinatorial chemistry, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0662] [Example 1] Concept of Accelerating Antigen Elimination Effect of Calcium-Dependent Antigen-Binding Antibody
[0663] (1-1) Antigen elimination acceleration effect of antigen-binding antibody based on pH-dependent binding antibody
[0664] H54 / L28-IgG1 described in WO 2009 / 125825 is a humanized anti-IL-6 receptor antibody, and Fv4-IgG1 binds to soluble human IL-6 receptor in a pH-dependent manner to H54 / L28-IgG1 A humanized anti-IL-6 receptor antibody made of the characteristics (binding under neutral conditions and dissociation under acidic conditions). In the mouse in vivo test described in WO 2009 / 125825, compared with the group administered with a mixture of H54 / L28-IgG1 and antigen-soluble human IL-6 receptor, Fv4-IgG1 and antigen-soluble human IL-6 receptor were administered It was shown in the group of mixtures of IL-6 receptors that the elimination of soluble human IL-6 receptors can be greatly accelerated.
[0665] The soluble human IL-6 receptor bound to t...
Embodiment 2
[0672] [Example 2] Obtaining a Ca-dependent binding antibody from a human antibody library using phage display technology
[0673] (2-1) Preparation of naive human antibody phage display library
[0674] Multiple human antibodies displaying Fab domains containing human antibody sequences were constructed according to (Methods Mol Biol. 2002;178:87-100.) using polyA RNA prepared from human PBMC or commercially available human polyA RNA as a template Phage display library.
[0675] (2-2) Obtaining Ca-dependent binding antibody fragments from a library by bead panning
[0676] Initial selection from the constructed human antibody phage display library was carried out by enriching only antibody fragments capable of binding to the antigen or by enriching them using Ca-dependent binding ability as an index. When concentrating antibody fragments having Ca-dependent binding ability, phages are eluted by chelating Ca ions with EDTA after binding to antigens in the presence of Ca ...
Embodiment 4
[0710] [Example 4] Calcium ion binding evaluation of the obtained antibody
[0711] Next, in order to evaluate the calcium ion binding of the antibody, the intermediate temperature of thermal denaturation (Tm value) was evaluated by differential scanning calorimetry (DSC) (MicroCal VP-Capillary DSC, manufactured by MicroCal). The intermediate temperature of thermal denaturation (Tm value) is an indicator of stability. When calcium ions are bound to stabilize the protein, the intermediate temperature of thermal denaturation (Tm value) is higher than when calcium ions are not bound (J Bio Chem. 2008 Sep 12 ; Vol.283 ; No. 37:pp 25140 - 25149), using this principle, the calcium ion binding of antibodies was evaluated. Dilute the purified antibody against 20 mM Tris-HCl, 150 mM NaCl, 2 mM CaCl 2 , pH7.4, or 20 mM Tris-HCl, 150 mM NaCl, 3 μM CaCl 2 , pH7.4 solution was dialyzed (EasySEP, TOMY). The protein solution was prepared at 0.1 mg / mL from the solution used for dialysis, a...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 