Monoclonal antibodies (mAbs) are important tools for modern biomedical research and treatment. Since Kohler and Milstein first used hybridoma technology to prepare monoclonal antibodies in 1975, the field has developed rapidly, not only playing a huge role in scientific research, but also showing broad application prospects in disease diagnosis and treatment. This article will comprehensively introduce the basic concepts, preparation processes, application fields, and future development of monoclonal antibodies.

1. Basic concepts of monoclonal antibodies

Monoclonal antibodies are highly uniform antibodies produced by a single B cell clone, targeting only specific antigenic epitopes. These antibodies have high specificity and homogeneity, and can accurately recognize and bind to target antigens, thereby exerting corresponding biological functions. The birth of monoclonal antibodies stems from a profound understanding of antibody diversity. There are about 2 × 10 ^ 12 lymphocytes in the adult body, of which T cells and B cells each account for about half. B cells are further subdivided into multiple clones, each with a similar structure but slightly different functions. Antigen determinants can specifically activate one or more cloned B cells, causing them to produce corresponding antibodies. These antibodies produced by a single B cell clone are called monoclonal antibodies.

The preparation of monoclonal antibodies mainly uses hybridoma technology. This technology forms a B cell hybridoma by fusing sensitized B cells with the ability to secrete specific antibodies and myeloma cells with unlimited proliferation capacity. This type of hybridoma cell not only has the characteristic of unlimited proliferation of tumor cells in vitro, but also has the characteristic of synthesizing and secreting antibodies to form cell specific antibodies. By culturing or intraperitoneal inoculation of mice, a large amount, high concentration, and very uniform antibody can be obtained. However, the Beacon Optofluidic System utilizes cutting-edge microfluidic and optoelectronic localization technology to achieve precise screening and cultivation of hybridoma cells. This system significantly shortens the antibody development cycle, taking only about 35 days from immunization to obtaining specific monoclonal antibodies, providing strong support for the effective progress of drug development and biological therapy.

2. Preparation process of monoclonal antibodies

1. Immune animals: The first step in preparing monoclonal antibodies is to immunize animals. Usually, female BALB/c mice aged 6-8 weeks are selected as immunization subjects and vaccinated according to a predetermined immunization regimen. Antigens enter peripheral immune organs through blood circulation or lymphatic circulation, stimulating the activation, proliferation, and differentiation of corresponding B lymphocyte clones, forming sensitized B lymphocytes.

2. Cell fusion: Extract mouse spleen, prepare spleen cell suspension, mix it with the prepared homologous myeloma cells in a certain proportion, and add fusion promoter polyethylene glycol. Under the action of polyethylene glycol, various lymphocytes fuse with myeloma cells to form hybridoma cells. Only fused hybridoma cells can survive and proliferate through selective medium selection, such as HAT medium.

3. Screening and cloning: Only a few hybridoma cells grown in HAT medium secrete specific monoclonal antibodies. Therefore, screening and cloning culture are necessary. Cloning culture usually uses limited dilution method to screen positive hybridoma cells that can produce the required monoclonal antibodies through sensitive, rapid, and specific immunological methods, and then perform cloning and amplification.

4. Large scale preparation of antibodies: The large-scale preparation of monoclonal antibodies mainly adopts animal in vivo induction method and in vitro culture method. The in vivo induction method involves injecting hybridoma cells into the abdominal cavity of mice, causing them to proliferate and produce and secrete monoclonal antibodies. The in vitro culture method includes culturing hybridoma cells in culture bottles and collecting monoclonal antibodies from the culture supernatant.

3. Application fields of monoclonal antibodies

1. Tumor treatment: Monoclonal antibodies play an important role in tumor treatment. They can be used alone as anti-tumor drugs or in combination with other chemotherapy drugs. By binding to specific antigens on the surface of tumor cells, monoclonal antibodies can block the growth and diffusion pathways of tumor cells, induce tumor cell apoptosis, or enhance the immune system's killing effect on tumor cells. For example, monoclonal antibody drugs have been used to treat non-small cell lung cancer, cancer, and other malignant tumors, achieving significant clinical efficacy.

2. Treatment of autoimmune diseases: Monoclonal antibodies have also shown broad application prospects in the treatment of autoimmune diseases. For example, in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus, monoclonal antibodies can alleviate the condition by inhibiting the activation of immune cells and the release of inflammatory factors. These antibodies are typically used in combination with traditional immunosuppressants to enhance therapeutic efficacy and reduce side effects.

3. Diagnosis and treatment of infectious diseases: In the field of infectious diseases, monoclonal antibodies are widely used for rapid detection and specific treatment of pathogens. By identifying and binding specific antigens on the surface of pathogens, monoclonal antibodies can accurately diagnose diseases in the early stages, providing strong support for timely intervention and treatment. In addition, some monoclonal antibodies have also developed into therapeutic antibodies that directly target pathogens or their toxins, such as antibody therapy against HIV, hepatitis B, hepatitis C and other viruses, and neutralizing antibodies against bacterial toxins (such as botulinum toxin and tetanus toxin).

4. Biomarker detection: Biomarkers are objective and measurable molecular indicators generated during the occurrence and development of diseases. Monoclonal antibodies play an important role in biomarker detection due to their high specificity and sensitivity. By constructing detection methods based on monoclonal antibodies, such as enzyme-linked immunosorbent assay (ELISA), flow cytometry, etc., precise detection of disease-related proteins, hormones, cytokines, and other biomarkers can be achieved, providing scientific basis for early diagnosis, disease monitoring, and prognosis evaluation of diseases.

5. Immunological research: Monoclonal antibodies play an irreplaceable role in immunological research. They are used to study the differentiation, activation, apoptosis, and other processes of immune cells, as well as the interaction mechanisms between immune molecules. By preparing monoclonal antibodies targeting specific immune cells or molecules, researchers can track the distribution, changes, and functional status of these cells or molecules in the body, thereby revealing the complexity and diversity of the immune system. In addition, monoclonal antibodies have been used to prepare immunotoxins, immunoadsorbents, and other tools, further promoting the in-depth development of immunological research.

4. Challenges and Future Prospects

Although monoclonal antibodies have achieved great success in the biomedical field, their development still faces some challenges. Firstly, in the preparation process, it is necessary to screen a large number of hybridoma cells to obtain monoclonal antibodies with high specificity and affinity, which is both time-consuming and laborious. Secondly, the immunogenicity of monoclonal antibodies in the human body limits their long-term use. To overcome these challenges, scientists are constantly exploring new preparation techniques and improvement strategies, such as genetically engineered antibodies, humanized antibodies, chimeric antibodies, etc.

In the future, with the continuous development of cutting-edge technologies such as gene editing, synthetic biology, and artificial intelligence, the preparation and application of monoclonal antibodies will face new opportunities and challenges. On the one hand, these technologies will greatly improve the preparation efficiency and specificity of monoclonal antibodies, reduce production costs and immunogenicity; On the other hand, it will also promote the expansion and deepening of the application of monoclonal antibodies in more disease fields. With the emergence of the Beacon Optofluidic System from Redbert (Beijing) Biotechnology Co., Ltd., it saves you a lot of time and greatly reduces production costs. The Beacon Optofluidic System can intervene when the cell diversity and survival rate reach their optimum after transfection, and it is easy to screen multiple cells and select cell lines with higher expression levels, thereby greatly reducing subsequent production costs. The emergence of Beacon Optofluidic System and monoclonal antibodies are expected to play a greater role in treating more diseases. We have reason to believe that in the near future, monoclonal antibodies will play a more important role in the biomedical field, contributing more wisdom and strength to the cause of human health.

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