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Venoms and Toxins

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ISSN (Print): 2666-1217
ISSN (Online): 2666-1225

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Research Article

Development and Validation of an ELISA to Evaluate Neutralizing Equine Anti Shiga Toxin Antibodies in Preclinical Studies

Author(s): Gonzalo Santiago, Hiriart Yanina, Luciana Muñoz, Florencia Rey, Gustavo Hein, Santiago Sanguineti, Vanesa Zylverman, Hugo H. Ortega* and Belkis E. Marelli*

Volume 2, Issue 2, 2022

Published on: 09 June, 2022

Article ID: e270422204096 Pages: 11

DOI: 10.2174/2666121702666220427081107

Open Access Journals Promotions 2
Abstract

Background: Hemolytic uremic syndrome associated with Shiga-toxin produced by Escherichia coli is a serious worldwide foodborne disease. Nowadays, no treatment is available, only supportive care can be provided, and 50 % of the patients require a period of dialysis. Recently, a therapy based on Neutralizing Equine Anti Shiga Toxin (NEAST) antibodies has been developed. NEAST is composed of F(ab’)2 fragments from equine immunoglobulins.

Objective: The purpose of this study was to develop an ELISA to measure serum concentrations of NEAST in mice and rabbits, and to validate it according to international recommendations. The validated method was further used to analyze the NEAST PK during preclinical studies.

Methods: A sandwich ELISA was developed, the performance of the calibration curve was assessed, and it was validated based on the parameters as accuracy, precision, specificity, selectivity, stability of the analyte, and dilutional linearity.

Results: This immunoassay was specific, sensitive, accurate and precise in a dynamic range from 7.81 to 500 ng/mL and from 15.63 to 500 ng/mL for mice and rabbits, respectively. This method was successfully applied to PK studies of NEAST after intravenous administration.

Conclusion: The results obtained are expected for a robust ELISA used for macromolecule analysis. Since NEAST is an equine F(ab′)2, this immunoassay would serve for the evaluation of the PK profile of any biological product based on molecules with similar characteristics. This immunoassay may be useful for current and future preclinical trials conducted for registration purposes.

Keywords: NEAST, ELISA, validation, preclinical studies, pharmacokinetic studies, toxin.

Graphical Abstract

[1]
Rosales A, Hofer J, Zimmerhackl L-B, et al. German-Austrian HUS Study Group. Need for long-term follow-up in enterohemorrhagic Escherichia coli-associated hemolytic uremic syndrome due to late-emerging sequelae. Clin Infect Dis 2012; 54(10): 1413-21.
[http://dx.doi.org/10.1093/cid/cis196] [PMID: 22412065]
[2]
Mostafavi A, Fozi MA, Koshkooieh AE, Mohammadabadi M, Babenko OI, Klopenko NI. Effect of LCORL gene polymorphism on body size traits in horse populations. Acta Sci Anim Sci 2019; 42: e47483.
[http://dx.doi.org/10.4025/actascianimsci.v42i1.47483]
[3]
Moazemi I, Mohammadabadi MR, Mostafavi A, et al. Polymorphism of DMRT3 gene and its association with body measurements in horse breeds. Russ J Genet 2020; 56(10): 1232-40.
[http://dx.doi.org/10.1134/S1022795420100087]
[4]
Asadollahpour N, Nosrati M, Mohammadabadi M. Genetic structure analysis of akhal-teke horse population and comparison with other horse breeds by using whole genome sequencing data. MGJ 2021; 16: 299-307.
[5]
Lang J, Attanath P, Quiambao B, et al. Evaluation of the safety, immunogenicity, and pharmacokinetic profile of a new, highly purified, heat-treated equine rabies immunoglobulin, administered either alone or in association with a purified, Vero-cell rabies vaccine. Acta Trop 1998; 70(3): 317-33.
[http://dx.doi.org/10.1016/S0001-706X(98)00038-2] [PMID: 9777717]
[6]
Quiambao BP, Dytioco HZ, Dizon RM, Crisostomo ME, Laot TM, Teuwen DE. Rabies post-exposure prophylaxis in the Philippines: Health status of patients having received purified equine F(ab’)(2) fragment rabies immunoglobulin (Favirab). PLoS Negl Trop Dis 2008; 2(5): e243.
[http://dx.doi.org/10.1371/journal.pntd.0000243] [PMID: 18509475]
[7]
Herbreteau CH, Jacquot F, Rith S, et al. Specific polyclonal F(ab’)2 neutralize a large panel of highly pathogenic avian influenza A viruses (H5N1) and control infection in mice. Immunotherapy 2014; 6(6): 699-708.
[http://dx.doi.org/10.2217/imt.14.40] [PMID: 24673720]
[8]
Bal C, Herbreteau CH, Buchy P, et al. Safety, potential efficacy, and pharmacokinetics of specific polyclonal immunoglobulin F(ab’)₂ fragments against avian influenza A (H5N1) in healthy volunteers: A single-centre, randomised, double-blind, placebo-controlled, phase 1 study. Lancet Infect Dis 2015; 15(3): 285-92.
[http://dx.doi.org/10.1016/S1473-3099(14)71072-2] [PMID: 25662592]
[9]
Lopardo G, Belloso WH, Nannini E, et al. INM005 Study Group. RBD-specific polyclonal F(ab´)2 fragments of equine antibodies in patients with moderate to severe COVID-19 disease: A randomized, multicenter, double-blind, placebo-controlled, adaptive phase 2/3 clinical trial. EClinicalMedicine 2021; 34: 100843.
[http://dx.doi.org/10.1016/j.eclinm.2021.100843] [PMID: 33870149]
[10]
Salinas F, Marelli BE, Sanguineti S, et al. Non-clinical safety assessment and in vivo biodistribution of CoviFab, an RBD-specific F(ab’)2 fragment derived from equine polyclonal antibodies. Toxicol Appl Pharmacol 2022; 434: 115796.
[http://dx.doi.org/10.1016/j.taap.2021.115796] [PMID: 34785274]
[11]
Chippaux JP, Lang J, Amadi-Eddine S, Fagot P, Le Mener V. Short report: Treatment of snake envenomations by a new polyvalent antivenom composed of highly purified F(ab)2: Results of a clinical trial in northern Cameroon. Am J Trop Med Hyg 1999; 61(6): 1017-8.
[http://dx.doi.org/10.4269/ajtmh.1999.61.1017] [PMID: 10674688]
[12]
Chippaux J-P, Massougbodji A, Stock RP, Alagon A. Investigators of African Antivipmyn in Benin. Clinical trial of an F(ab’)2 polyvalent equine antivenom for African snake bites in Benin. Am J Trop Med Hyg 2007; 77(3): 538-46.
[http://dx.doi.org/10.4269/ajtmh.2007.77.538] [PMID: 17827375]
[13]
Hiriart Y, Pardo R, Bukata L, et al. Preclinical studies of NEAST (Neutralizing Equine Anti-Shiga To Xin): A potential treatment for prevention of stec-hus. Int J Drug Dev Res 2019; 11: 15-24.
[14]
Karliner JS, Belaval GS. Incidence of reactions following administration of antirabies serum. JAMA 1965; 193(5): 359-62.
[http://dx.doi.org/10.1001/jama.1965.03090050035009] [PMID: 14313890]
[15]
de Haro L, Lang J, Bedry R, et al. Envenimations par vipères européennes. Etude multicentrique de tolérance du Viperfav, nouvel antivenin par voie intraveineuse. Ann Fr Anesth Reanim 1998; 17(7): 681-7.
[http://dx.doi.org/10.1016/S0750-7658(98)80105-6] [PMID: 9750806]
[16]
Marcato P, Mulvey G, Read RJ, et al. Immunoprophylactic potential of cloned Shiga toxin 2 B subunit. J Infect Dis 2001; 183(3): 435-43.
[http://dx.doi.org/10.1086/318080] [PMID: 11133375]
[17]
Boyer L, Degan J, Ruha AM, Mallie J, Mangin E, Alagón A. Safety of intravenous equine F(ab’)2: Insights following clinical trials involving 1534 recipients of scorpion antivenom. Toxicon 2013; 76: 386-93.
[http://dx.doi.org/10.1016/j.toxicon.2013.07.017] [PMID: 23916602]
[18]
Darwish IA. Immunoassay methods and their applications in pharmaceutical analysis: Basic methodology and recent advances. Int J Biomed Sci 2006; 2(3): 217-35.
[PMID: 23674985]
[19]
European medicines agency, committee for medicinal products for human use, . Guideline on Bioanalytical Method Validation. In: London, UK 2012. Available from: https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-bioanalytical-method-validation_en.pdf
[20]
U.S Food and Drug Administration, U.S. Department of Health and Human Services. Guidance for industry: Bioanalytical method validation. Rockville, USA 2018. Available from: https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf
[21]
Organisation for Economic Co-operation and Development (OECD). Principles on good laboratory practice. In: Paris, France 1998. Available from: http://www.oecd.org/officialdocuments/publicdisplaydocumentpdf/?cote=env/mc/chem(98)17&doclanguage=en
[22]
Laplagne DA, Zylberman V, Ainciart N, et al. Engineering of a polymeric bacterial protein as a scaffold for the multiple display of peptides. Proteins 2004; 57(4): 820-8.
[http://dx.doi.org/10.1002/prot.20248] [PMID: 15390265]
[23]
Mejias MP, Ghersi G, Craig PO, et al. Immunization with a chimera consisting of the B subunit of Shiga toxin type 2 and brucella lumazine synthase confers total protection against Shiga toxins in mice. J Immunol 2013; 191(5): 2403-11.
[http://dx.doi.org/10.4049/jimmunol.1300999] [PMID: 23918978]
[24]
Suessenbach FK, Tins J, Burckhardt BB. LENA Consortium. Customisation and validation of a low-volume plasma renin activity immunoassay: Enabling of regulatory compliant determination in paediatric trials. Pract Lab Med 2019; 17: e00144.
[http://dx.doi.org/10.1016/j.plabm.2019.e00144] [PMID: 31867426]
[25]
Maple L, Lathrop R, Bozich S, et al. Development and validation of ELISA for herceptin detection in human serum. J Immunol Methods 2004; 295(1-2): 169-82.
[http://dx.doi.org/10.1016/j.jim.2004.09.012] [PMID: 15627622]
[26]
NRC. Guide for the Care and Use of Laboratory Animals. National Academies Press 2011.
[27]
Kleinstreuer NC, Tong W, Tetko IV. Computational toxicology. Chem Res Toxicol 2020; 33(3): 687-8.
[http://dx.doi.org/10.1021/acs.chemrestox.0c00070] [PMID: 32172570]
[28]
European Medicines Agency, Committee for Medicinal Products for Human Use. Guideline on the Investigation of Bioequivalence. 2010. Available from: https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-investigation-bioequivalence-rev1_en.pdf
[29]
Reagan-Shaw S, Nihal M, Ahmad N. Dose translation from animal to human studies revisited. FASEB J 2008; 22(3): 659-61.
[http://dx.doi.org/10.1096/fj.07-9574LSF] [PMID: 17942826]
[30]
Lee JW, Devanarayan V, Barrett YC, et al. Fit-for-purpose method development and validation for successful biomarker measurement. Pharm Res 2006; 23(2): 312-28.
[http://dx.doi.org/10.1007/s11095-005-9045-3] [PMID: 16397743]
[31]
Mander A, Chowdhury F, Low L, Ottensmeier CH. Fit for purpose? A case study: Validation of immunological endpoint assays for the detection of cellular and humoral responses to anti-tumour DNA fusion vaccines. Cancer Immunol Immunother 2009; 58(5): 789-800.
[http://dx.doi.org/10.1007/s00262-008-0633-z] [PMID: 19066888]
[32]
Chowdhury F, Tutt AL, Chan C, Glennie M, Johnson PW. Development, validation and application of ELISAs for pharmacokinetic and HACA assessment of a chimeric anti-CD40 monoclonal antibody in human serum. J Immunol Methods 2010; 363(1): 1-8.
[http://dx.doi.org/10.1016/j.jim.2010.09.023] [PMID: 20869964]
[33]
Findlay JW, Smith WC, Lee JW, et al. Validation of immunoassays for bioanalysis: A pharmaceutical industry perspective. J Pharm Biomed Anal 2000; 21(6): 1249-73.
[http://dx.doi.org/10.1016/S0731-7085(99)00244-7] [PMID: 10708409]
[34]
DeSilva B, Smith W, Weiner R, et al. Recommendations for the bioanalytical method validation of ligand-binding assays to support pharmacokinetic assessments of macromolecules. Pharm Res 2003; 20(11): 1885-900.
[http://dx.doi.org/10.1023/B:PHAM.0000003390.51761.3d] [PMID: 14661937]
[35]
Smolec J, DeSilva B, Smith W, et al. Bioanalytical method validation for macromolecules in support of pharmacokinetic studies. Pharm Res 2005; 22(9): 1425-31.
[http://dx.doi.org/10.1007/s11095-005-5917-9] [PMID: 16132353]
[36]
Hampson G, Ward TH, Cummings J, et al. Validation of an ELISA for the determination of rituximab pharmacokinetics in clinical trials subjects. J Immunol Methods 2010; 360(1-2): 30-8.
[http://dx.doi.org/10.1016/j.jim.2010.05.009] [PMID: 20547164]
[37]
Cummings J, Ward TH, Greystoke A, Ranson M, Dive C. Biomarker method validation in anticancer drug development. Br J Pharmacol 2008; 153(4): 646-56.
[http://dx.doi.org/10.1038/sj.bjp.0707441] [PMID: 17876307]
[38]
Miller KJ, Bowsher RR, Celniker A, et al. Workshop on bioanalytical methods validation for macromolecules: Summary report. Pharm Res 2001; 18(9): 1373-83.
[http://dx.doi.org/10.1023/A:1013062600566] [PMID: 11683255]
[39]
Lobo ED, Hansen RJ, Balthasar JP. Antibody pharmacokinetics and pharmacodynamics. J Pharm Sci 2004; 93(11): 2645-68.
[http://dx.doi.org/10.1002/jps.20178] [PMID: 15389672]
[40]
Quesada L, Sevcik C, Lomonte B, Rojas E, Gutiérrez JM. Pharmacokinetics of whole IgG equine antivenom: Comparison between normal and envenomed rabbits. Toxicon 2006; 48(3): 255-63.
[http://dx.doi.org/10.1016/j.toxicon.2006.05.010] [PMID: 16863656]
[41]
Vázquez H, Chávez-Haro A, García-Ubbelohde W, Paniagua-Solís J, Alagón A, Sevcik C. Pharmacokinetics of a F(ab’)2 scorpion antivenom administered intramuscularly in healthy human volunteers. Int Immunopharmacol 2010; 10(11): 1318-24.
[http://dx.doi.org/10.1016/j.intimp.2010.08.018] [PMID: 20849955]
[42]
Vázquez H, Olvera F, Alagón A, Sevcik C. Production of anti-horse antibodies induced by IgG, F(ab’)2 and Fab applied repeatedly to rabbits. Effect on antivenom pharmacokinetics. Toxicon 2013; 76: 362-9.
[http://dx.doi.org/10.1016/j.toxicon.2013.09.004] [PMID: 24047962]

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