Antibody engineering, as an important component of modern biotechnology, processes and recombines antibody genes through recombinant DNA and protein engineering techniques to create novel antibody molecules with specific functions. These antibody molecules not only retain the specificity and primary biological activity of natural antibodies, but also demonstrate broader application prospects by removing or replacing unrelated structures. This article will delve into the basic principles, development history, technical methods, application status, and future trends of antibody engineering.

1、Basic principles of antibody engineering

Antibodies, also known as immunoglobulins (Ig), are natural biomolecules produced by plasma cells or stimulated memory B cells. They have a Y-shaped heterodimer structure, consisting of two 25kDa light chains and two at least 50kDa heavy chains, connected by multiple disulfide bonds and non covalent interactions. Antibodies can be divided into antigen binding domains (Fab) and crystallizable fragment domains (Fc). The Fab region is responsible for binding to antigens, while the Fc region is involved in various immune effector functions such as antibody dependent cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), and antibody dependent cell phagocytosis.

The core of antibody engineering lies in the use of recombinant DNA and protein engineering techniques to modify antibody genes. These technologies include gene cloning, gene editing, gene expression regulation, etc., aimed at optimizing the specificity, affinity, stability, and production efficiency of antibodies. Through genetic engineering technology, novel antibody molecules with specific functions can be designed, such as bispecific antibodies, antibody drug conjugates (ADCs), etc.

2、The Development History of Antibody Engineering

1. Early antibody preparation technology: Antibodies have a history of over 100 years as preparations for disease prevention, diagnosis, and treatment. The early methods of preparing antibodies mainly involved immunizing animals with a certain natural antigen and extracting polyclonal antibodies from animal serum. However, the heterogeneity of polyclonal antibodies limits their further research and application.

2. The birth of monoclonal antibody technology: In 1975, Kohler and Milstein first used B lymphocyte hybridoma technology to prepare homogeneous monoclonal antibodies (mAbs). The birth of this technology is considered the first qualitative leap in the development of antibody engineering and a milestone in the development of modern biotechnology. Monoclonal antibodies have been widely used in disease diagnosis, treatment, and scientific research, but issues such as mouse origin, high production costs, and poor ability to penetrate blood vessel walls still need to be addressed.

3. Development of genetic engineering antibody technology: In the early 1980s, the research results on the structure and function of antibody genes were combined with recombinant DNA technology to produce the technology of genetic engineering antibodies. Genetic engineering antibodies are processed, modified, and recombined with antibody genes through genetic engineering techniques, and then introduced into appropriate receptor cells for expression. Compared with monoclonal antibodies, genetically engineered antibodies have the advantages of reducing human rejection reactions, small molecular weight, strong penetration ability, and low production costs.

3、Antibody engineering technology methods

1. Gene cloning and editing: Gene cloning is the foundation of antibody engineering, which involves obtaining antibody gene sequences through techniques such as PCR and gene synthesis, and cloning them into expression vectors. Gene editing techniques such as CRISPR/Cas9 are used for site directed mutagenesis, insertion, or deletion of antibody genes to optimize antibody performance.

2. Expression system: The commonly used expression systems in antibody engineering include prokaryotic cells (such as E. coli), eukaryotic cells (such as yeast, insect cells, and mammalian cells), and plant cells. Different expression systems have their own advantages and disadvantages, such as the low cost and high yield of E. coli expression systems, but there are issues with endotoxins and insufficient post-translational modifications; Mammalian cell expression systems can undergo complex post-translational modifications, but at a higher cost.

3. Antibody modification techniques: Antibody modification techniques include humanization, affinity maturation, Fc region modification, etc. Humanization technology reduces human rejection of antibodies by replacing mouse derived sequences with human derived sequences. Affinity maturation technology enhances the affinity of antibody molecules through high-frequency somatic mutations and screening. Fc region modification is used to optimize the immune response function of antibodies, such as prolonging half-life, enhancing ADCC and CDC effects, etc.

4、The current application status of antibody engineering

1. Biopharmaceutical field: Antibody engineering plays an important role in the biopharmaceutical field and has become an important means of treating diseases such as cancer and autoimmune diseases. At present, a variety of antibody based therapeutic drugs have been approved for marketing by the U.S. Food and Drug Administration (FDA), such as trastuzumab for breast cancer and tozumab for rheumatoid arthritis.

2. Infectious disease prevention and control: Antibody engineering also plays an important role in infectious disease prevention and control. By preparing neutralizing antibodies against specific pathogens, viral infection and transmission can be effectively blocked. For example, the neutralizing antibody sotrovimab against SARS CoV-2 has been approved by FDA for emergency use authorization (EUA), providing a new option for the treatment of COVID-19.

3. Research and Diagnosis: Antibody engineering is also widely used in the fields of research and diagnosis. By preparing specific antibodies, detection and localization of specific proteins, cells, or tissues can be achieved, providing powerful tools for scientific research. Meanwhile, antibodies are also one of the important biomarkers for disease diagnosis, such as tumor markers, autoimmune disease markers, etc.

5、The future development trend of antibody engineering

1. The continuous emergence of new antibody technologies: With the continuous development of biotechnology, new antibody technologies continue to emerge. For example, bispecific antibodies can simultaneously bind to two different antigens or different epitopes of the same antigen, which has a wider range of application prospects. Antibody drug conjugates (ADCs) couple antibodies with cytotoxic drugs to achieve precise targeting of tumor cells. In addition, new types of antibodies such as nanobodies and antibody mimetics have also shown great potential.

2. Cross border integration and innovation of antibody engineering: Antibody engineering is currently undergoing cross-border integration and innovation with other fields. For example, the combination of artificial intelligence and big data technologies can accelerate the screening and optimization process of antibodies; Combining synthetic biology and metabolic engineering can optimize the production process of antibodies and reduce costs. These cross-border integrations and innovations will inject new impetus into the development of antibody engineering.

3. Social impact and sustainable development of antibody engineering: The application of antibody engineering in disease treatment, biopharmaceuticals and other fields not only improves people's quality of life and health level, but also promotes the development of related industries and economic growth. Meanwhile, the application of antibody engineering in environmental protection, resource utilization, and other aspects also contributes to achieving sustainable development goals. For example, environmental remediation can be achieved by preparing antibodies against pollutants; Reduce production costs and resource consumption by optimizing the production process.

Antibody engineering, as an important component of modern biotechnology, processes and recombines antibody genes through recombinant DNA and protein engineering techniques to create novel antibody molecules with specific functions. These antibody molecules play important roles in fields such as biopharmaceuticals, infectious disease prevention and control, scientific research, and diagnosis. With the launch of the Beacon Optofluidic System by Redbert (Beijing) Biotechnology Co., Ltd., it can save you a lot of screening time and greatly reduce production costs. The conventional use of hybridoma or phage display technology usually takes 3-6 months, but the Beacon Optofluidic System device can obtain specific antibody sequences in just 3 days. Single plasma cells can be directly isolated and detected in a 0.5nl system, from which target cells expressing specific antibodies can be screened, and their heavy and light chain mRNAs can be obtained. These mRNAs can be directly used for sequencing and optimization after reverse transcription. We believe that antibody engineering will continue to play an important role in disease treatment, diagnosis, and prevention in the future, making greater contributions to human health.

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