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. Oak Biosciences has 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 ChainLight Chain
HumanIgA
IgD
IgG1, IgG2, IgG3, IgG4
IgM
IgE
Igκ, Igλ2
MouseIgA
IgD
IgG1, IgG2a, Ig2b, IgG3
IgM
IgE
Igκ, Igλ1, Igλ2
RabbitIgA
IgG
IgM
IgE
Igκ1, Igκ2, Igλ
RatIgA
IgD
IgG1, IgG2a, IgG2b, IgG2c
IgM
IgE
IgG2b, IgM
Igκ, Igλ
ChickenIgY, IgMIgλ

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 FcFcR/C1q BindingEffector Function
1human IgG1 (P257I/Q311I)Increased binding to FcRnUnchanged half-life
2human IgG1 (N297Q)Compromised or ablated FcγRI, FcγRII, FcγRIII and the C1q
Non-glycosylated antibody
3human IgG1 (S239D/I332E/G236A)Increased FcγRIIa/FcγRIIb ratioIncreased macrophage phagocytosis
4human IgG1 (K326W/E333S)
Increased binding to C1qIncreased CDC
5human IgG1 (S239D/I332E)Increased binding to FcγRIIIa
Increased ADCC
6human IgG1 (S239D/A330L/I332E)Increased binding to FcγRIIIa
Increased ADCC
7human IgG1 (E333A)Increased binding to FcγRIIIaIncreased ADCC and CDC
8human IgG1 (E233P/L234V/L235A/ΔG236 +A327G/A330S/P331S)Reduced binding to FcγRIReduced ADCC and CDC
9human IgG1 (I253A)Decreased binding to FcRnDecreased half-life
10human IgG1 (M252Y/S254T/T256E)Increased binding to FcRnIncreased half-life
11human IgG1 (M428L/N434A)Increased binding to FcRnIncreased half-life
12human IgG1 (M252Y/S254T/T256E +H433K/N434F)Increased binding to FcRnIncreased half-life
13human IgG1 (T250Q/M428L)Increased binding to FcRnIncreased 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 FcFcR/C1q BindingEffector Function
human IgG2 (K322A)Reduced binding to C1qReduced 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 FcFcR/C1q BindingEffector Function
human IgG4 (S228P)N/AReduce 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 FcFcR/C1q BindingEffector Function
1mouse IgG2a(L235E+ E318A/K320A/K322A)Reduced binding to FcγRI and C1qReduced ADCC and CDC
2mouse IgG2a (N297Q)Compromised or ablated FcγRI, FcγRII, FcγRIII, and the C1qNon-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

CatalogDescriptionCommentsPrice
84101Molecular construction of IgG expression vector; Transient expression; Purification by protein AIgG from rabbit, pig, dog, catContact Us
84102Molecular construction of IgG expression vector; Transient expression; Purification by protein GIgG from mouse, human, cow, goat and sheepContact US
84103Molecular construction of IgG expression vector; Transient expression; Purification by protein A/GIgG from many speciesContact US
84104Molecular construction of expression vector; Transient expression; Purification by protein LAll classes of immunoglobulin (i.e., IgG, IgM, IgA, IgE and IgD), possessing the appropriate Igk light chains Contact Us

 

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Chimeric Antibody Production and Affinity Purification Services