Introduction: Pharmaceutical companies as well as food and cosmetics manufacturers are legally required to provide a shelf-life label on their products packaging as part of their stability study report. There are different recommended software like R software package and SAS which can perform as shelf-life estimating tools for analyzing the data achieved by the stability testing of drugs and vaccines. Methods: Recombinant hepatitis B surface antigen vaccine (Pasteur Institute of Iran) was used as a sample for the entire quality control assays according to Pharmacopeia and NIH (National Institute of Health) procedures. For the stability study, full test examinations were done and R software package was applied to estimate the shelf-life. Results: The results of R software indicated a variety of statistical information which makes the data interpretation more intelligible and apprehensible. Conclusion: Based on our results and experience, the best way to obtain a shelf-life or the expiration date is to calculate it manually however, using software such as R can increase the accuracy of the results.

Introduction: Polyclonal antibodies are required to be affinity purified. Improved purification methods of polyclonal antibody provide an opportunity to pick the most purified immunoglobulins as a primary or secondary antibody in immunoassays that are included in many vaccine studies. Two common techniques for purifying proteins is salt precipitation and chromatography purification. Our work focuses on purification of polyclonal antibodies against recombinant human erythropoietin (EPO) antigen using these techniques. Methods: A polyclonal antibody was produced by antigen injection with Freund's adjuvant into female albino rabbits. After separation of immunoglobulins using caprylic acid and ammonium sulfate precipitation, selected samples were analyzed by ion exchange chromatography for separation of polyclonal antibody from albumin. The purified proteins were analyzed by SDS-PAGE and antibody was detected by Western Blot analysis and ELISA. Results of immunodiffusion test detected polyclonal antibody production in rabbits. Results: Caprylic acid precipitation was shown to be a more effective purification method than ammonium sulfate. Analysis of protein by spectrophotometer showed 97.6% purity by caprylic acid and 77% purity by ammonium sulfate method. Western Blot and ELISA tests confirmed the presence of antibody against EPO. Conclusion: These findings suggest that caprylic acid can be used as a quality control method in a production facility with minimal cost. On the other hand, ion exchange chromatography is the most common purification method for proteins. Therefore, combination of these techniques may effectively reduce contaminations in antibody purification procedures which may positively affect the interpretations of vaccine efficacies.

Introduction: Endotoxin removal is a crucial stage in ensuring the safety of parenteral products. S3E3-S-Sepharose, which has been generated via site-specific immobilization of the S3E3 cationic amphiphilic peptide (CAP) on Sepharose, is a newly designed affinity matrix proposed for lipopolysaccharide (LPS) removal from biopharmaceuticals and vaccines. Methods: In the current study, the kinetic behavior of LPS adsorption on the matrix was investigated at pHs 4.5 and 8.5 by incubation of LPS-contaminated bovine serum albumin (BSA) solutions, as a model, with the S3E3-Sepharose matrix at different incubation times in a batch-wise mode. Various mathematic models were employed to explain the amount of adsorbed LPS, and the normalized root mean square error (NRMSE), relative prediction error (RE), and relative percentage error (RPE) were utilized to identify the best-fitting model. Results: The kinetics study revealed that the pseudo-second-order (PSO) reaction, and pore diffusion mass transfer were the rate-controlling steps of LPS adsorption on the S3E3-S-Sepharose and pH of samples did not affect the LPS adsorption kinetics. Conclusion: These findings provide valuable insights for scaling up the LPS removal process through affinity chromatography, contributing to advances in biopharmaceutical and vaccine production research.