Discussions

Antibody and protein production is a fundamental aspect of biotechnology and pharmaceuticals, involving the synthesis of these crucial biomolecules for research, therapeutic, and industrial purposes.

Antibody Production
Design and Selection: The process begins with identifying the specific antigen against which antibodies are to be produced. Monoclonal antibodies are typically generated using hybridoma technology or recombinant DNA techniques, while polyclonal antibodies are produced in animals.

Expression Systems: Antibodies can be produced in various expression systems, including mammalian cells (e.g., CHO or HEK293), which offer post-translational modifications similar to those in humans, ensuring functionality and stability.

Purification: The antibodies are then purified using affinity chromatography, exploiting their specific binding capabilities to isolate them from other proteins and impurities.

Characterization: Following purification, antibodies undergo rigorous characterization to confirm their specificity, affinity, and purity, using techniques such as ELISA, Western blotting, and surface plasmon resonance (SPR).

Protein Production
Gene Cloning: The gene encoding the protein of interest is cloned into an expression vector. This step may involve optimizing the gene sequence for the host system to enhance expression levels.

Optimization of Expression Systems: Proteins are produced in various systems such as bacteria (E. coli), yeast, insect cells, or mammalian cells, depending on the required post-translational modifications and yield.

Cultivation and Induction: The host cells are cultivated under optimal conditions, and expression of the protein is induced. Conditions such as temperature, medium composition, and induction time can significantly influence yield and activity.

Purification and Validation: Post-expression, proteins are purified through chromatography techniques like ion exchange, size exclusion, and affinity purification. Structural and functional validation ensures that the protein is correctly folded and active.