Computational Engineering of Protein L to Achieve an Optimal Affinity Chromatography Resin for Purification of Antibody Fragments

• Published in: Analytical chemistry (Washington) Vol. 93; no. 46; pp. 15253 - 15261
• Main Authors: Rahmati, Saman, Torkashvand, Fatemeh, Amanlou, Massoud, Bagherzadeh, Kowsar, Fard Esfahani, Pezhman, Aghamirza Moghim Aliabadi, Hooman, Vaziri, Behrouz
• Format: Journal Article
• Language: English
• Published: Washington American Chemical Society 23.11.2021
• Subjects: Affinity; Affinity chromatography; Analytical chemistry; Antibodies; Binding; Chains; Chemistry; Chromatography; Computer applications; Fragments; Ligands; Light chains; Molecular docking; Molecular dynamics; Protein engineering; Protein L; Protein purification; Proteins; Purification; Purity; Recovery; Resins
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.1c01871

Protein L affinity chromatography is a useful method for the purification of antibody fragments containing kappa light chains. In affinity chromatography, increasing the binding affinity leads to increased product purity, recovery, and dynamic binding capacity (DBC). In this study, molecular docking and molecular dynamics simulation techniques were used to design the engineered Protein L with higher affinity to the kappa light chain. Each engineered ligand was produced as a recombinant protein and coupled to a solid matrix. The purity, recovery, and DBC of the engineered resins were evaluated and then compared to those of a commercially available resin. The results showed important parameters for engineering more efficient Protein L ligands for affinity chromatography.