Bispecific Antibody Expression Service

Alpha Lifetech has developed a number of methods to produce bispecific antibodies. We can produce various forms of bispecific antibodies, provide various antibody purification services, and offer a variety of expression systems for customers to choose from. We provide one-stop bispecific antibody expression services, from gene synthesis, vector construction and optimization, bispecific antibody expression and purification, antibody final validation and delivery, with strict quality control at every step to meet customer needs.

Introduction to bispecific antibody

Bispecific antibodies (bsAb) refer to two different binding domains, which can simultaneously bind to two antigens or two different epitopes of the same antigen. The specificity and multi-target binding related characteristics of bsAb make it another major progress after monoclonal antibody therapy. Bispecific antibodies are mainly used in cancer treatment, and a small number are also used in autoimmune diseases, chronic inflammation, hematological diseases, etc. As of the end of 2023, 14 bsAbs have been approved: 11 for cancer treatment and 3 for non oncology indications.

Bispecific antibodies come in many forms, ranging, for example, from small proteins consisting of only two antigen-binding fragments in tandem to IGG-like immunoglobulins containing additional domains.

Over the past two decades, bispecific and multispecific antibodies have made significant progress in the field of tumor therapy. These antibodies are able to bind two or more antigens at the same time, improving treatment accuracy and reducing side effects by accurately distinguishing malignant cells from normal cells.

Fig 1: The development of bispecific antibody. Reference source: Li, Heliang, 2020. Challenges and strategies for next-generation bispecific antibody-based antitumor therapeutics.

Structure of bispecific antibodies

IgG-like Antibody

The molecular weight of IgG-like is relatively large, which is helpful for purification, has high stability and solubility, a long serum half-life, and high biological activity. It retains the corresponding function of Fc fragments, but the corresponding tissue permeability is low, making production difficult. These structures mainly include Triomab, Knobs into holes, Double Variable Domain Immunoglobulin (DVD Ig), scFv2 Fc, etc.

Non-IgG-like Antibody

Non-IgG-like antibodies do not include the Fc region, resulting in significant structural differences from natural antibodies. They have a short plasma half-life, poor stability, and are prone to polymerization and inactivation. However, they are relatively easy to produce, avoiding chain cross-linking issues. Moreover, by expressing two polypeptide chains simultaneously in eukaryotic or prokaryotic expression systems, bispecific antibodies with this structure can be produced without the need for chemical coupling or assembly steps, thereby increasing yield and reducing production costs. These structures mainly include BiTE, bispecific T-cell engager， TandAb, tandem diabody， DART, dual affinity retargeting.

Fig2: Representative bispecific antibodies and their format. Reference source: Sun, Yanze, 2023. Bispecific antibodies in cancer therapy: Target selection and regulatory requirements.

Bispecific Antibody Production Process

bispecific antibody production-Alpha Lifetech

Fig3: Bispecific antibodies production process

Bispecific Antibodies Analysis Service Workflow

Steps	Service Content	Timeline
Gene Synthesis	Generation of up to 3 different design schemes of sequences, de novo generation of recombinant antibody DNA plasmids	2-3 weeks
Small Scale Production	Production of bispecific antibody proteins → Small scale expression in mammalian cell lines → Validation by SDS-PAGE → Binding analysis with recombinant protein antigens by ELISA → Comparison of clones with parental antibodies	5-6 weeks
Antibody Identify	Expression of the first two full-length bispecific antibodies → Binding analysis by ELISA → ELISA evaluation of antibodies recognized by anti mouse antibodies	1 week
Antibody Production	Large scale bispecific antibody expression	3-4 weeks

Our Advantages

We cover the upstream and downstream services of antibody production, and can provide technical services from antibody preparation, bispecific antibody purification and antibody separation and purification, antibody sequencing, antibody validation, etc, , which can be used for bispecific antibody therapy.

High Expression

High protein expression in CHO cells.
Stable Storage

>2 weeks in serum at 37 °C.
Related Services

Bispecific antibody development, bispecific antibody purification, antibody drug conjugation, etc
High Success Rates

Multiple antibody forms can be produced with high success rates and multiple purification methods available
No Solubility Issue

>30 mg/ml
Stable Cell Lnes

Many stable cell lines with tumor antigens have been developed to validate many bispecific antibodies