Proposed reduction of the in vivo pyrogen test by the in vitro LAL assay for the quality control of anticrotallic, antiscorpion, antirabies and antitetanus sera

• Published in: Toxicology in vitro 2019-09, Vol.59, p.292-299
• Main Authors: Fingola, Fernando F., Albertino, Sheila R.G., Abrantes, Shirley de M.P., Zamith, Helena P.S.
• Format: Article
• Language: English
• Subjects: Animals; Antitoxins - analysis; Assaying; Correlation coefficients; Crotalid Venoms - immunology; CV; Endotoxin; Endotoxin limit concentration; Endotoxins; Health surveillance; In vitro methods and tests; In vivo methods and tests; LAL chromogenic kinetic assay; Method reduction; Parameters; Pyrogen; Pyrogens; Quality Control; Rabies virus - immunology; Recovery; Regulatory agencies; Scorpion Venoms - immunology; Sensitivity analysis; Surveillance; Tetanus Toxin - immunology
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2019.04.031

The rationale for a formal study of the LAL (kinetic chromogenic method) assay for anticrotallic (SAC), antirabies (SAR), antitetanus (SAT) and antiscorpion (SAE) sera involved the determination of parameters required by the Brazilian National Health Surveillance Agency (ANVISA), the USA Food and Drug Administration (FDA), USP (United States Pharmacopeia, 39) and ICH (International Conference on Harmonization). The curve correlation coefficients obtained with the standard endotoxin control ranged from −0.980 to −1.000 in all experiments. Endotoxin recovery added to the SAC, SAR, SAT and SAE samples, at the working dilutions (1:10, 1:10, 1:10 and 1, 100 respectively), met the criteria required by the FDA, USP and Brazilian ANVISA for the Inhibition-Potentiation test. The applied methodology for the four analyzed sera fulfilled the required criteria for all performance parameters. Thus, the present study demonstrated that the in vivo pyrogen test can be potentially replaced by the LAL assay for all assessed sera samples displaying higher sensitivity and following the 3 Rs principle, in addition to maintaining quality control in Sanitary Surveillance. •A rationale for a formal study of reduction of the in vivo pyrogen test (PT) by the LAL assay was performed.•The LAL assay was carried out for anticrotallic, antirabies, antitetanus and antiscorpion sera.•Parameters required by Brazilian and international agencies were determined.•Methodology assessment for the four sera fulfilled required criteria for all performance parameters.•The LAL assay can replace the PT for the assessed sera, with higher sensitivity and following the 3 Rs principle.