3 . 2023

Development of a Pharmacopoeia Standard for the quantitative determination of the activity of human immunoglobulin antirhesus Rho(D)

Abstract

Introduction. The pharmacological effect of human immunoglobulin (HI) antirhesus Rh_o(D) drugs is characterized by their specific activity – the ability of anti-D antibodies (Ab) IgG to selectively block an undesirable immune response to the D antigen of Rh-positive fetal erythrocytes that have entered the mother’s bloodstream, as well as the dose of the administered. The dose of the drug is calculated based on the quantitative content of anti-D-IgG-Ab, expressed in International Units (IU) or in micrograms (mcg). Foreign manufacturers the content of anti-D-IgG-Ab in preparations determined quantitatively by hemagglutination, flow cytofluorimetry and enzyme immunoassay in IU or mcg (5 IU = 1 mcg) in relation to the International Standard (IS) of anti-D immunoglobulin or a standard sample (SS) calibrated relative to the IS. Domestic manufacturers determined the content of anti-D-IgG-Ab in the titers using an indirect Coombs method without using a SS. The absence of a national SS certified in IU (or mcg) did not allow domestic manufacturers of HI anti-rhesus Rh_o(D) drugs to establish the norm of anti-D-IgG-Ab content in the drug in IU (or mcg) to calculate the dose of administration in accordance with international requirements for its clinical use, which could cause the absence of the desired therapeutic effect, erythrocyte hemolysis or the development of other adverse reactions.

The aim – to develop a Pharmacopoeia Standard (PhS) for the quantitative determination of the activity of HI antirhesus Rh_o(D).

Material and methods. Samples of the candidate for PhS, IS anti-D immunoglobulin, SS of the European Pharmacopoeia with the content of anti-D-IgG-Ab determined in the agglutination reaction (1:8), human erythrocytes of the phenotypes R1R1 and R2R2, rr were used in study. To establish the certified characteristics of the PhS, the enzyme immunoassay and the hemagglutination in gel were used.

Results. The national PhS 3.1.000452 of the State Pharmacopoeia of the Russian Federation «Standard sample for determining the activity of IH antirhesus Rh_o(D) (content of anti-D-IgG-Ab)» has been developed with an activity value of 516 IU/ml (103 mcg/ml) and an extended uncertainty: ±100 IU/ml (±20 mcg/ml) (coverage ratio 2, confidence level 95 %). It has been established that the available validated enzyme immunoassay using erythrocytes of the R1R1 or R2R2 phenotypes for self-preparation of an immunosorbent provides an accurate quantitative determination of anti-D-IgG-Ab in the PhS. It is shown that the use of enzyme immunoassay and hemagglutination in gel to establish the certified characteristics of the PhS ensures the traceability and comparability of the results obtained.

Conclusion. The developed PhS is included in the register of PhS of biological origin, as well as implemented in the regulatory documentation for the drug produced in the Russian Federation (INN «Human Immunoglobulin anti-rhesus Rh_o(D)»). The use of PhS makes it possible to establish the values of the anti-D-IgG-Ab content in the drug in IU/ml (or mcg/ml), which contributes to an adequate calculation of the dose of administration during its clinical use for specific perinatal prevention of Rh-immunization of women with Rh-negative blood affiliation.

Keywords:national pharmacopoeia standard; human immunoglobulin antirhesus Rho(D); quantitative determination; anti-D-antibodies IgG

For citation: Shvedova E.V., Klepikova A.G., Fadeikina O.V., Shvedov D.V., Borisevich I.V., Soldatov A.A. Development of a Pharmacopoeia Standard for the quantitative determination of the activity of human immunoglobulin antirhesus Rh_o(D). Immunologiya. 2023; 44 (3): 358–67. DOI: https://doi.org/10.33029/1816-2134-2023-44-3-358-367 (in Russian)

Funding. The study was carried out as part of a publicly funded research project No. 056-00052-23-00 and was supported by the Scientific Centre for Expert Evaluation of Medicinal Products of the Ministry of Health of the Russian Federation (R&D public accounting No. 121022000147-4).

Conflict of interests. The authors declare no conflict of interests.

Authors’ contribution. Research concept and design – Shvedova E.V.; experimental work – Shvedova E.V., Klepikova A.G.; analysis of experimental data – Fadeikina O.V.; writing a text – Shvedova E.V.; general management – Shvedov D.V.; final approval of the version for publication – Borisevich I.V., Soldatov A.A.

References

1. Savel’eva G.M., Kurcer M.A., Panina O.B., Sichinava L.G., Klimenko P.A., Konopljannikov A.G., Alekseenkova M.V. Diagnosis, treatment, prevention of fetal hemolytic disease in RH-sensitization. Rossiyskiy Vestnik Perinatologii i Pediatrii (Russian Bulletin of perinatology and pediatrics). 2006; 6: 73–8. (in Russian)

2. Olovnikova N.I., Nikolaeva T.L., Ershler M.A. Immunization of rhesus immunization: the prospect of a monoclonal immunoglobulin to prevent haemolytic disease of the newborn. Immunologiya. 2018; 39. (1): 74–80. DOI: http://doi.org/10.18821/0506-4952-2018-39-1-74-80 (in Russian)

3. Manylova N.A., Loginova I.I. Using the Kleihauer–Betke test in revealing of fetomaternal transfusion and determination of risk factors of anemia in newborns of the first week of life. System analysis and management in biomedical systems. 2008; 7 (1): 142–4. (in Russian)

4. Guideline on the clinical investigation of human anti-D immunoglobulin for intravenous and/or intramuscular use: EMA/CPMP/BPWG/575/99, rev. 1. URL: http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500003334.pdf

5. Shvedova E.V., Kudasheva E.Yu., Klimov V.I. Methods for evaluating the specific activity of preparations of human antirhesus Rh_o(D): current status of the problem. Immunologiya. 2020; 41 (3): 256–1. DOI: http://doi.org/10.33029/0206-4952-2020-41-3-256-261 (in Russian)

6. Vildanova N.S., Kormshchikova E.S., Kalinina E.N. Human anti-D immunoglobulin preparations: potency standardisation milestones. BIOpreparaty. Profilaktika, diagnostika, lechenie. 2022; 22 (3): 241–8. DOI: https://doi.org/10.30895/2221-996X-2022-22-3-241-248 (in Russian)

7. Shvedova E.V., Kudasheva E.Yu., Leshina S.A., Davydov D.S. Method of quantitative determination of IgG anti-D antibodies in human immunoglobulin preparations Rh_o(D) antirhesus. Patent for invention № 2777845 dated 11.08.2022, Russian Federation, WIPO: А61К35/12,15; G01N33/53; applicant and patent holder SCEEMP of the MOH of Russia, application № 2021137704 dated 20.12.2021 (in Russian)

8. Tests for anti-D antibodies in human immunoglobulin drugs. GPM.1.8.2.0004.15 State Pharmacopoeia of the Russian Federation XIVed. URL: https://femb.ru/record/pharmacopea14 (in Russian)

9. Glantz S. Biomedical statistics. Moscow; Praktika, 1999: 459 р. (in Russian)

10. ICH guidelines for the pharmaceutical industry. Quality. In: Beregovyh V.V. (ed.). Saint Petersburg: Professiya, 2017: 768 p. (in Russian)

11. 2.7.13. Assay of Human Anti-D Immunoglobulin 01/2008:20713. European Pharmacopoeia, 11^th ed. Strasboung: Directorate for the Quality of Medicines of the Council of Europe.

12. Trusov G.A., Chaplenko A.A., Semenova I.S. Use of flow cytometry to quality evaluation of biomedical cell products. Biological Products. Prevention, Diagnosis, Treatment. 2018; 18 (1): 16–4. DOI: https://doi.org/10.30895/2221-996X-2018-18-1-16-24 (in Russian)

13. Shvedova E.V., Leshina S.A., Davydov D.S., Borisevich I.V., Kudasheva E.Yu. Development of criteria for immobilization of human erythrocytes of phenotypes R1R1 and R2R2 on the solid phase in determining the content of anti-D-antibodies IgG in immunoglobulin human antirhesus Rh_o(D) preparations by enzyme immunoassay. Immunologiya. 2022; 43 (2): 208–16. DOI: https://doi.org/10.33029/0206-4952-2022-43-2-208-216 (in Russian)

14. WHO International Standard Anti-D Immunoglobulin. NIBSC code: 16/332. Instructions for use. URL: https://www.nibsc.org/documents/ifu/16-332.pdf

15. Register of Pharmacopoeia reference samples of the State Pharmacopoeia of the Russian Federation. URL: http://www.regmed.ru/produkt-n-service/fso/fso-registry/fso-of-biological-origin

16. State Register of Medicines of the Ministry of Health of the Russian Federation. URL: http://grls.rosminzdrav.ru/grls.aspx (in Russian)

17. Accuracy (trueness and precision) of measurement methods and results. Part 1. General principles and definitions. 5725-1-2002. URL: https://docs.cntd.ru/document/1200029975 (in Russian)

18. State system for ensuring the uniformity of measurements. Reference materials of materials (substances). Basic principles. 8.753-2011. URL: https://docs.cntd.ru/document/1200093434 (in Russian)

19. Shvedova E.V., Leshina S.A., Kudasheva E.Yu., Borisevich I.V., Shvedov D.V., Merkulov V.A. Development of acceptance criteria for evaluating the results of quantitative determination of anti-D-antibodies IgG in immunoglobulin human antirhesus Rh_o(D) preparations by enzyme immunoassay. Biological Products. Prevention, Diagnosis, Treatment. 2022; 22 (3): 266–8. DOI: https://doi.org/10.30895/2221-996X-2022-22-3-266-278 (in Russian)

20. Iskakova O.A. Gel technologies for the immune hematological blood testing. Science and Healthcare. 2013; 1: 48–2. (in Russian)

21. Bioanalytical method validation guidance for industry. U.S. Biopharmaceutics, May 2018. URL: http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htm

22. EDQM Immunoglobulin panel for anti-D antibodies test BRP. Instructions for use. URL: https://crs.edqm.eu/db/4DCGI/db/4DCGI/leaflet?leaflet=Y0000540_1.pdf