PRODUCTION OF MONOCLONAL ANTIBODIES.pptx

PRODUCTION OF MONOCLONAL ANTIBODIES

1. PRODUCTION OF MONOCLONAL ANTIBODIES P:3 U:2 Vedanti S. Gharat Roll No. – 09 M.Sc. Biotechnology Part- 2

2. INTRODUCTIO N Monoclonal Antibody - A type of protein that is made in the laboratory and can bind to certain targets in the body, such as antigens on the surface of cancer cells . A monoclonal antibody (mAb) is originally produced by a single B-cell . Production of mAbs was first introduced in 1975 using cell fusion technique and hybridoma cell production. A Hybridoma is formed by fusion of an antibody producing B-lymphocyte and a myeloma cell line.

3. These cells have two main characteristics, production of uniform, monospecific antibodies (mAbs) that originate from the B-cell and immortality that comes from the myeloma cell line. A hybridoma cell line is thus acting like a biological factory that produces and secretes mAbs into the cell culture medium. Most of the mAbs have been produced in mice and are thus proteins of murine origin . mAbs were later found to be able to bind biological targets like tumor antigens, molecules involved in autoimmune and infectious disease-related molecules,  etc . This led to emergence of therapeutic monoclonal antibodies.

4. The most common type of mAb used to treat cancer are naked monoclonal antibodies . Examples of naked mAb include alemtuzumab (Campath, Genzyme) for the treatment of chronic lymphocytic leukemia, and trastuzumab (Herceptin, Genentech) for the treatment of stomach and breast cancers that contain the HER-2 protein . Conjugated monoclonal antibodies — also known as labeled, loaded or tagged antibodies. Examples of conjugated mAb include ibritumomab tiuxetan (Zevalin, Spectrum), which is used to treat certain types of non-Hodgkin’s lymphoma, and ado-trastuzumab emtansine (Kadcyla, Genentech), for the treatment of HER-2–positive breast cancer . Other mAb are bispecific . An example of a bispecific mAb is blinatumomab (Blincyto, Amgen), used to treat certain types of acute lymphocytic leukemia.

5. PRODUCTION OF mAb The traditional mAb production process usually starts with generation of mAb-producing cells (i.e. hybridomas) by fusing myeloma cells with desired antibody-producing splenocytes (e.g. B cells ). These B cells are typically sourced from animals. After cell fusion, large numbers of clones are screened and selected on the basis of antigen specificity and immunoglobulin class. Once candidate  hybridoma cell lines  are identified, each "hit" is confirmed, validated, and characterized using a variety of downstream functional assays . Upon completion, the clones are scaled up where additional downstream bioprocesses occur.

6. A TYPICAL mAb PRODUCTION PROCESS Immunization of mice & isolation of splenocytes Preparation of myeloma cells Fusion Clone screening and picking Functional characterization Scale up and wean Expansion

7. Several critical advances in unit operations for the production of monoclonal antibodies (MAbs) reported within the last 20 years, with a focus on novel quasi‐continuous downstream purification schemes/ platforms being explored in academic and industrial settings. The primary drivers for these advances are based on five central ideals : ( i) D ecreased production costs (ii) Equivalent or (ideally) improved product quality/uniformity (iii) Increased flexibility of processing equipment and facilities ( iv) Elimination of unnecessary process/hold steps ( v) Reduction of plant facilities footprint.

8. ADVANTAGES OF mAb Same quality of the antibody is maintained amongst the different production batches. Highly reproducible and scalable, unlimited production source. Speed and sensitivity and specificity of assays. Can produce antibodies when needed. No need to worry about maintaining the animals. Antigen or immunogen need not be pure. Selection helps to identify the right clones against the specific antigen.

9. REFERENCES What are monoclonal antibodies? https ://www.healio.com/news/hematology-oncology/20151104/what-are-monoclonal-antibodies#:~: text=Examples%20of%20naked%20monoclonal%20antibodies,contain%20the%20HER%2D2%20protein . Therapeutic Monoclonal Antibodies and Emergence of Their Biosimilars https :// www.ncbi.nlm.nih.gov/pmc/articles/PMC5960060/ Producing Monoclonal Antibodies https :// www.sinobiological.com/resource/antibody-technical/monoclonal-antibody-production Monoclonal Antibody Production https://www.moleculardevices.com/applications/monoclonal-antibody- production#:~:text=The%20traditional%20monoclonal%20antibody%20(mAb ,sourced%20from%20animals%2C%20usually%20mice . https :// www.pdfdrive.com/biosimilars-of-monoclonal-antibodies-a-practical-guide-to-manufacturing-preclinical-and-clinical-development-e188662797.html Book.

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