CD19 and CD20 bispecific CAR targeting diagram

Bispecific Antibody Expression

Antibody Production
Home > Antibody Production > Bispecific Antibody Expression

Our bispecific antibody production will provide you high-quality bispecific antibodies in 3 to 4 weeks.

Biointron supports the expression of all bispecific formats including but not limited to: Knobs-into-holes IgG, IgG-scFv fusion, CrossMab, Y-body, Bispecific T-cell Engagers (BiTEs), and more. We utilize mammalian cell systems, such as CHO-K1 and HEK293 for antibody expression, ensuring high yield and quality. You will receive SDS-PAGE, SEC-HPLC, and endotoxin results along with the purified bispecific antibody and expression plasmids containing your gene of interest.

Biointron offers multiple platforms for antibody discovery services to generate candidate monoclonal antibodies, which can then be engineered into bispecific formats tailored to your research or therapeutic goals.

Our recombinant antibody production services also include:

Scientists monitoring bispecific antibody production in a cleanroom
Hexavalent multispecific IgG with Fab and VHH domains on the Fc region
Multispecific IgG with tandem variable domains on both Fab arms
Asymmetric bispecific IgG with an additional VHH domain
Multispecific IgG with VHH domains fused to Fab and Fc regions
Multispecific IgG with additional variable domains on both Fab arms
Multispecific IgG with additional VH and VL domains on both Fab arms
Asymmetric bispecific IgG with two distinct antigen-binding arms
Dual-variable-domain bispecific IgG with tandem VH and VL domains
VHH and extracellular-domain fusion for multispecific antibody binding
Linear Fab-VHH-extracellular-domain multispecific antibody format
Linear multispecific antibody with tandem variable domains and an extracellular domain
Bivalent VHH fusion linked through an extracellular domain
Linear multispecific antibody with tandem Fab and VHH domains
Linear trispecific antibody with Fab, VHH and extracellular domain components
Linear multispecific antibody combining Fab, VHH and extracellular domains
Linear multispecific antibody with tandem Fab, VHH and extracellular domains
Multispecific IgG with extracellular domains fused to the Fc region
IgG scaffold with four VHH domains for multispecific antibody binding
Bispecific IgG with tandem VHH domains on each Fab arm
Multispecific IgG with extracellular domains fused to Fab and Fc termini
Multispecific antibody combining Fab, VHH and extracellular domains
Multispecific IgG with VHH and extracellular domains fused to an IgG scaffold
Tetravalent multispecific IgG with VHH domains on Fab and Fc termini
Tetravalent multispecific antibody with VHH domains fused to an IgG scaffold

Some examples showcasing the various formats of Biointron's previous multispecific antibody productions.

Advantages of Our Bispecific Antibody Production Service

Customized Service

  • Any format can be expressed
  • Cross verify structural design

Large Capacity

  • Automated working stations
  • Multiple methods for purification and QC

Strong Expertise

  • 2-3 weeks (from sequences to BsAbs)
  • Over forty thousand of bispecific antibodies have been delivered

Bispecific Antibody Production for Research Success with Biointron

The ever-increasing research and clinical demands call for advanced therapeutic antibody drugs, and bispecific antibodies are among the most promising. lmmunoglobulin G (lgG), the most common antibody type in blood circulation, is typically monospecific and recognizes only a single antigen, with the exception of lgG4. In contrast, bispecific antibodies are designed to recognize and bind to two different targets simultaneously, providing enhanced or unique effects through distinct mechanisms.

Certified to ISO 9001:2015 and supported by a global team of 600+ professionals, Biointron offers a fully integrated, high-throughput solution - from gene sequence to purified antibody - in just 2 weeks.

Bispecific antibodies have demonstrated their therapeutic value in clinical settings, as they can be designed to redirect immune cells (such as T cells) toward diseased cells for targeted elimination. Additionally, bispecific antibodies can facilitate the delivery of therapeutic payloads, such as toxins or drugs, to in vivo targets.

Custom Bispecific Antibodies Produced by Biointron

With over 12 years of experience in antibody discovery, expression, and optimization, Biointron has become a trusted partner to more than 3,000 biotech and pharmaceutical companies worldwide. Since 2012, we’ve delivered over 100,000 recombinant antibodies annually.

Choosing the optimal bispecific antibody format and structure is a critical decision influenced by application, manufacturing considerations, and commercial goals. Our expert team is here to guide you through the selection process and support your project every step of the way.

Our Streamlined Bispecific Antibody Production Process

Gene synthesis

Plasmid construction and preparation

Transient expression

Affinity purification

Stringent quality control

Fast delivery

Bispecific Antibody Expression Service Details

Our bispecific antibody production service follows a comprehensive workflow to ensure quality and efficiency at every step.

Milestone Service Description Timeline Deliverables
Gene synthesis & cloning
  • Codon optimization and gene synthesis
  • Subcloning into an expression vector
  • Plasmid amplification and preparation
1 week
  • CoA report
  • Expression plasmid contain GOI
  • Purified antibody
Transient expression & purification
  • Co-Transfection of mammalian cells
  • Transient expression and purification
  • QC analysis (SDS-PAGE, SEC-HPLC, A280, Endotoxin)
1-2 weeks
  • Gene synthesis & cloning
    Timeline

    1 week

    Description
    • Codon optimization and gene synthesis
    • Subcloning into an expression vector
    • Plasmid amplification and preparation
    Deliverables
    • CoA report
    • Expression plasmid contain GOI
    • Purified antibody
  • Transient expression & purification
    Timeline

    1-2 weeks

    Description
    • Co-Transfection of mammalian cells
    • Transient expression and purification
    • QC analysis (SDS-PAGE, SEC-HPLC, A280, Endotoxin)
    Deliverables
    • CoA report
    • Expression plasmid contain GOI
    • Purified antibody

Case Study

Case 1: 1-step purification of IgG-scFv

This case highlights the one-step purification of two IgG-scFv fusion constructs, with the scFv fused to either the N- or C-terminus of the IgG. Following 4-day expression, both constructs achieved high purity, demonstrating the efficiency of the one-step purification process.

bispecific-case1-1
bispecific-case1-2
bispecific-case1-2
Yield (mg/L) SDS-PAGE SEC (280 nm)
253.3 >95% 99.440%
bispecific-case1-3
bispecific-case1-2
bispecific-case1-2
Yield (mg/L) SDS-PAGE SEC (280 nm)
230.05 >95% 99.440%

Case 2:

bispecific-case1-1

This case involves the expression of an asymmetric multi-specific antibody with a molecular weight of 193 kDa. Following one-step Protein A purification, SDS-PAGE and analytical SEC-HPLC revealed the presence of half antibodies and hole-hole homodimers. Due to the close pI values, mixed-mode chromatography was employed for polishing, effectively removing most of the impurities.

Protein A purification

bispecific-case1-1

SEC-HPLC: 71.17%

Protein A purification

bispecific-case1-1

SEC-HPLC: 97.17%

Case 3:

bispecific-case1-1

This case involves the expression of a CrossMabVH-VL. After one-step Protein A purification, the ratio of the target heterodimer was difficult to determine. Mass spectrometry analysis revealed impurities such as half antibodies and constructs containing identical light chains. To further purify the product, hydrophobic interaction chromatography (HIC) was applied, successfully removing most of the impurities.

bispecific-case1-1
bispecific-case1-1

Target heterodimer: 65%
By-products: HC1+LC1; HC1+HC2+2*LC1

2nd purification: HIC
bispecific-case1-1
bispecific-case1-1

Target heterodimer: 96%

Team Member Spotlight

“We like to ensure our clients are more than satisfied with our bispecific antibodies, offering a wide range of customizable formats to suit your precise research needs. Bispecific antibodies are transforming the landscape of medicine.”
Jiasha Huang, Biointron bispecific antibody expression scientist
Jiasha Huang
Antibody Expression Team

FAQs About Bispecific Antibody Production

Resources

Services & Products

Our website uses cookies to improve your experience. Read our Privacy Policy to find out more.