JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY HYDERABAD B. Pharmacy III PHARMACEUTICAL BIOTECHNOLOGY Previous Question Paper (November/December-2020 - R17)

Protein engineering is the process of developing useful or valuable proteins by modifying their amino acid sequences through genetic manipulation. This field aims to alter the structure, stability, catalytic activity, or binding affinity of proteins to suit specific industrial, medical, or research applications.

The primary methods for engineering proteins include:

• Site-directed mutagenesis: A precise technique used to introduce specific point mutations at a predetermined location in the DNA sequence to study or change the function of a single amino acid residue.
• Random mutagenesis: Involves creating a large library of protein variants by introducing random mutations across the gene, often using error-prone PCR or chemical mutagens, followed by screening for desired traits.
• DNA shuffling: A method of directed evolution where fragments of homologous genes are recombined to create chimeric proteins with improved properties.
• Rational design: Uses computational modeling and structural biology data to predict the effect of specific mutations on protein folding and function before experimental validation.
• De novo design: The creation of entirely new protein sequences and structures that do not exist in nature, designed from scratch to perform specific functions.

Two examples of proteins used in diagnostics and therapeutics are:

• Insulin: A therapeutic protein used for the treatment of diabetes mellitus to regulate blood glucose levels.
• Monoclonal Antibodies (e.g., Trastuzumab): Used extensively in both diagnostics (to detect specific antigens in immunoassays like ELISA) and therapeutics (to target cancer cells or modulate the immune system).