Trivalent trispecific antibody constructs
A specific and antigenic technology, applied in the direction of antibodies, specific peptides, anti-tumor drugs, etc., can solve the problems of poor expression and folding of antibody chains, poor stability of assembled antibodies, complex in vitro assembly reactions or purification methods, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0425] 6.13.2. Example 1: Bivalent monospecific and bivalent bispecific constructs
[0426] A bivalent monovalent monovalent recognizing TNFα was constructed using standard molecular biology procedures following the following architecture (VL(certolizumab)-CH3(knob)-CH2-CH3 / VH(certolizumab)-CH3(hole)) Specific B-Body. In this construct,
[0427] First polypeptide chain (SEQ ID NO: 1)
[0428] Domain A = VL (certolizumab)
[0429] Domain B = CH3 (IgG1) (Knob: S354C+T366W)
[0430]Domain D = CH2 (IgG1)
[0431] Domain E = CH3 (IgG1)
[0432] Second polypeptide chain (SEQ ID NO:2)
[0433] Domain F = VH (certolizumab)
[0434] Domain G=CH3 (IgG1) (holes: Y349C, T366S, L368A, Y407V)
[0435] The third polypeptide chain:
[0436] Same as the first polypeptide chain
[0437] The fourth polypeptide chain:
[0438] Same as the second polypeptide chain.
[0439] Domain and polypeptide chain references conform to image 3 . The entire construct architecture is in Figure ...
Embodiment 2
[0448] 6.13.3. Example 2: Bivalent bispecific B-Body "BC1"
[0449] We constructed a bivalent bispecific construct specific for PD1 and a second antigen "Antigen A", which we named "BC1". The salient features of the "BC1" architecture are Image 6 shown in .
[0450] In more detail, the architecture (with image 3 The domain and polypeptide chain references and modifications of the native sequence indicated in brackets) are:
[0451] First polypeptide chain (SEQ ID NO:8)
[0452] Domain A=VL("Antigen A")
[0453] Domain B = CH3 (T366K; 445K, 446S, 447C tripeptide insertion)
[0454] Domain D=CH2
[0455] Domain E=CH3 (T366W, S354C)
[0456] Second polypeptide chain (SEQ ID NO:9):
[0457] Domain F = VH ("Antigen A")
[0458] Domain G=CH3 (L351D; 445G, 446E, 447C tripeptide insertion)
[0459] Third polypeptide chain (SEQ ID NO: 10):
[0460] Domain H = VL ("Nivo")
[0461] Domain I = CL(κ)
[0462] Domain J=CH2
[0463] Domain K=CH3 (Y349C, D356E, L358M, T366S, L...
Embodiment 6
[0534] 6.13.7. Embodiment 6: Variable CH3 connection engineering
[0535] We generated a series of variants in which we mutated the VL-CH3 linkage between domain A and domain B and the VH-CH3 linkage between domain F and domain G to evaluate bivalent 1x1 B-Body construction The expression level, assembly and stability of the body. Although many solutions are possible, in order to reduce the introduction of T-cell epitopes, we chose to use only residues found naturally within the VL, VH and CH3 domains. Structural evaluation of the domain architecture further constrains desired sequence combinations. Tables 2 and 3 below show linkages based on various linkage variants of "BC1" and other bivalent constructs.
[0536]
[0537]
[0538]
[0539]
[0540] Figure 20 shows the size exclusion chromatograms of "BC15" and "BC16" samples at 40°C for the indicated weeks of the accelerated stability testing protocol. "BC15" remained stable; "BC16" proved to be unstable over ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


