Recombinant Antibody Production and Affinity Purification
Recombinant antibodies or xhimeric antibodies or are antibodies produced by using recombinant antibody coding genes. They mostly consist of a heavy and light chain of the variable region of immunoglobulin. Chimeric antibodies have many advantages in both medical and research applications, which make them a popular subject of exploration and new production against specific targets. Fusion Biolabs have developed a unique technology for rapid production of full length chimeric antibodies, either native or engineered Fc.
We have manufactured about 150 recombinant antibodies using our CHO expression system yearly. The productivity is from 400 mg to 950 mg per liter. For IP issues, we could not deliver the expression vectors; however, we could swap them to any commercial or your own vectors. The Mab we manufactured using our procedure is audited by top pharmaceutic companies and is proved for in vivo animal use. The quality and quantity are guaranteed.
Recombinant antibodies contain variable domains, sequenced from your hybridoma cell line, which can be combined with any constant domains or engineered constant domains.
a) Recombinant Antibodies with Native Fc
Some of native Fc immunoglobulin listed in the following table:
Native Fc Immunoglobulin (Different Species, Classes and Isotypes)
| Heavy Chain | Light Chain | |
|---|---|---|
| Human | IgA IgD IgG1, IgG2, IgG3, IgG4 IgM IgE | Igκ, Igλ2 |
| Mouse | IgA IgD IgG1, IgG2a, Ig2b, IgG3 IgM IgE | Igκ, Igλ1, Igλ2 |
| Rabbit | IgA IgG IgM IgE | Igκ1, Igκ2, Igλ |
| Rat | IgA IgD IgG1, IgG2a, IgG2b, IgG2c IgM IgE IgG2b, IgM | Igκ, Igλ |
| Chicken | IgY, IgM | Igλ |
b) Recombinant Antibodies with Engineered Fc (human IgG1)
– Increased ADCC and no change CDC
– Increased binding to C1q and iIncreased CDC
– Increased binding to FcRn and unchanged half-life
– Increased binding to FcγRIIIa and increased ADCC
– Increased binding to FcγRIIIa and increased ADCC and CDC
– Reduced binding to FcγRI and reduced ADCC and CDC
– Increased binding to FcRn and increased half-life
– Decreased binding to FcRn and decreased half-life
Some of Human IgG1 Engineered Fc listed in the following table:
Engineered Isotypes of human IgG1
Engineered Fc FcR/C1q Binding Effector Function
1 human IgG1 (P257I/Q311I) Increased binding to FcRn Unchanged half-life
2 human IgG1 (N297Q) Compromised or ablated FcγRI, FcγRII, FcγRIII and the C1q
Non-glycosylated antibody
3 human IgG1 (S239D/I332E/G236A) Increased FcγRIIa/FcγRIIb ratio Increased macrophage phagocytosis
4 human IgG1 (K326W/E333S)
Increased binding to C1q Increased CDC
5 human IgG1 (S239D/I332E) Increased binding to FcγRIIIa
Increased ADCC
6 human IgG1 (S239D/A330L/I332E) Increased binding to FcγRIIIa
Increased ADCC
7 human IgG1 (E333A) Increased binding to FcγRIIIa Increased ADCC and CDC
8 human IgG1 (E233P/L234V/L235A/ΔG236 +A327G/A330S/P331S) Reduced binding to FcγRI Reduced ADCC and CDC
9 human IgG1 (I253A) Decreased binding to FcRn Decreased half-life
10 human IgG1 (M252Y/S254T/T256E) Increased binding to FcRn Increased half-life
11 human IgG1 (M428L/N434A) Increased binding to FcRn Increased half-life
12 human IgG1 (M252Y/S254T/T256E +H433K/N434F) Increased binding to FcRn Increased half-life
13 human IgG1 (T250Q/M428L) Increased binding to FcRn Increased half-life
c) Recombinant Antibodies with Engineered Fc (human IgG2)
– Reduced binding to C1q and reduced CDC
Some of Human IgG2 Engineered Fc listed in the following table:
Engineered Isotypes of human IgG2
| Engineered Fc | FcR/C1q Binding | Effector Function |
|---|---|---|
| human IgG2 (K322A) | Reduced binding to C1q | Reduced CDC |
d) Recombinant Antibodies with Engineered Fc (human IgG4)
– reduce Fab-arm exchange
Some of Human IgG4 Engineered Fc listed in the following table:
Engineered Isotypes of human IgG4
| Engineered Fc | FcR/C1q Binding | Effector Function |
|---|---|---|
| human IgG4 (S228P) | N/A | Reduce Fab-arm exchange |
e) Recombinant Antibodies with Engineered Fc (mouse IgG2a)
– Reduced binding to FcγRI and C1q and reduced ADCC and CDC
Some of mouse IgG2a Engineered Fc listed in the following table:
Engineered Isotypes of mouse IgG2a
| Engineered Fc | FcR/C1q Binding | Effector Function | |
|---|---|---|---|
| 1 | mouse IgG2a(L235E+ E318A/K320A/K322A) | Reduced binding to FcγRI and C1q | Reduced ADCC and CDC |
| 2 | mouse IgG2a (N297Q) | Compromised or ablated FcγRI, FcγRII, FcγRIII, and the C1q | Non-glycosylated antibody |
Our proprietary recombinant antibody technology allows you to specify your requirements by PURIFIED AMOUNT, NOT BY CULTURE VOLUME.
Our scientists have extensive experience in constructing and expressing recombinant or chimeric antibodies of various isotypes, including modified constant regions. We specializes in expression in CHO or HEK293 cells using our proprietary high mammalian expression systems.
Key Features
- Codon optimization technology
- Seamless cloning technology
- Choice of any species, isotypes and class
- Full line of engineered Fc (human IgG1, IgG2 and IgG4 and mouse IgG2a)
- Fast transient expression (1-2 weeks) with scale from 0.1 to 10 g
- Requested and paid by final purified AMOUNT (GUARANTEED quality and quantity), not by culture volume (estimated purified amount)
Process Overview
Client provides: Antibody sequence, plasmids containing the coding sequences, or hybridoma cells (>1×105 cells, in liquid nitrogen or dry ice) with isotype or class information
Stage I: Variable Region (VH and VL) Sequencing or Gene Synthesis
Stage II: Expression Vector Construction
Cloning into our above ready-to-use native or engineered Fc vector Constructionm (ready-to-use)
Stage III: Transient Expression and mAb Purification
Deliverables: 0.1 mg – 10 g of purified chimeric antibody, data sheet and full report
Stage IV: Cell Line Production (optional)
Turnaround time: 1- 2 weeks via transient expression; 6-10 weeks via cell line production.
Ordering Information
Chimeric Antibody or Recombinant Antibody production and Affinity Purification Services
Catalog Description Comments Price
84101 Molecular construction of IgG expression vector; Transient expression; Purification by protein A IgG from rabbit, pig, dog, cat Contact Us
84102 Molecular construction of IgG expression vector; Transient expression; Purification by protein G IgG from mouse, human, cow, goat and sheep Contact US
84103 Molecular construction of IgG expression vector; Transient expression; Purification by protein A/G IgG from many species Contact US
84104 Molecular construction of expression vector; Transient expression; Purification by protein L All classes of immunoglobulin (i.e., IgG, IgM, IgA, IgE and IgD), possessing the appropriate Igk light chains Contact Us
For complex projects, please fill the form below for a customized quotation and proposal!