Optimizing recombinant antibody production is important for cost-effective therapeutics and diagnostics. With downstream impact on commercialization, higher productivity is highly sought after beyond laboratory scales, where efficient production can also accelerate antibody characterizations and investigations. Using HEK293E cells as the base model for mammalian antibody production, various transfection and culture parameters were systematically analyzed using antibody light chain production before applying them onto whole antibody production. Transfection parameters investigated include seeding cell density, the concentration of the transfection reagent and DNA, complexation time, temperature, and volume, as well as culture parameters such as media replacement, serum deprivation, media volume, post-transfection harvest, incubation temperature, and common nutrient supplements. Incorporating the most optimized parameters, Pertuzumab κ-chain only and whole recombinant Pertuzumab antibody production were found to increase by 452 % and 252 % respectively, that can be used to guide future cost-effective transient antibody production with the potential for further scaling up.