EFFECT OF AMINO ACID ADDITION AS STABILIZERS ON TOTAL PROTEIN CONTENT OF INTRAVENOUS THERAPEUTIC HUMAN NORMAL IMMUNOGLOBULIN

MANOJ KUMAR RAJPUT; MANVANDRA PRATAP SINGH; APRAJITA SINGH; TARA CHAND; ANOOP KUMAR; MEENA KUMARI

doi:10.22159/ajpcr.2024v17i7.50708

Authors

MANOJ KUMAR RAJPUT Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India. https://orcid.org/0000-0002-3550-7333
MANVANDRA PRATAP SINGH Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India.
APRAJITA SINGH Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India.
TARA CHAND Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India.
ANOOP KUMAR Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India.
MEENA KUMARI Blood Products Laboratory, National Institute of Biologicals, Noida, Uttar Pradesh, India.

DOI:

https://doi.org/10.22159/ajpcr.2024v17i7.50708

Keywords:

Human immunoglobulin, Quality control, Protein estimation, Kjeldahl, Biuret

Abstract

Objectives: This study was conducted to show the variation in total protein content of therapeutic immunoglobulin batches after the addition of free amino acids.

Methods: A total of 10 different batches of therapeutic human normal immunoglobulin from five different manufacturers were taken. The total protein content in these samples was determined by both Kjeldahl and Biuret methods. The total protein in these samples was also determined by both methods after the addition of 0.3 M glycine, 0.3 M arginine, and 0.3 M histidine separately.

Results: In neat samples, both methods quantified similar total protein content. However, both methods showed different protein contents where amino acids such as glycine, arginine, or histidine have been added. The total protein content by the Kjeldahl method in the samples added with glycine, arginine, or histidine was found higher as compared to the Biuret method.

Conclusion: Our study suggests that the manufacturers of therapeutic immunoglobulin should replace amino acid stabilizers with nonnitrogenous stabilizer because the Kjeldahl method recommended in various pharmacopeias is based on the detection of total nitrogen. Therefore, the addition of any nitrogenous stabilizer will be quantified as protein and results may be out of specification when minimum and maximum limits are prescribed in the pharmacopeia.

Downloads

Download data is not yet available.

References

Stiehm ER. Human gamma globulins as therapeutic agents. Adv Pediatr. 1988;35:1-72. doi: 10.1016/S0065-3101(24)00173-7, PMID 2461056

AQ4 EMEA Committee for Proprietary Medicinal Products (CPMP): Note for guidance on plasma derived medicinal products. CPMP/ BPWG/269/95. Ser. 2. London, 1998. Core. SPC for human anti-D immunoglobulin and human anti-D immunoglobulin for intravenous use, CPMP/BPWG/574/99, London, 1999. Core SPC for human normal immunoglobulin for intravenous administration (IVIg), CPMP/ BPWG/859/95 rev. Vol. 1, London, 1999. Core. SPC for human normal immunoglobulin for subcutaneous and intramuscular use (SC/IMIg) CPMP/BPWG/282/00, London; 2002.

AQ4 Pharmacopoea Europaea. Europaisches Arzneibuch. Amtliche Deutsche Ausgabe, DAV Stuttgart; 1997. p. 1289.

Gaines A, Varricchio F, Kapit R. Renal insufficiency and failure associated with immune globulin intravenous therapy-United States, 1985-1998. MMWR Morb Mortal Wkly Rep. 1999;48(24):518-21. PMID 10401909

Patil V, Kaveri SV. The mechanisms of action of IVIG in autoimmune and inflammatory diseases. ISBT Sci Ser. 2013;8(1):185-8. doi: 10.1111/voxs.12037

Szenczi A, Kardos J, Medgyesi GA, Zaávodszky P. The effect of solvent environment on the conformation and stability of human polyclonal IgG in solution. Biologicals. 2006;34(1):5-14. doi: 10.1016/j. biologicals.2005.06.007, PMID 16168667

Thakral S, Sonje J, Munjal B, Suryanarayanan R. Stabilizers and their interaction with formulation components in frozen and freeze-dried protein formulations. Adv Drug Deliv Rev. 2021;173:1-19. doi: 10.1016/j.addr.2021.03.003, PMID 33741437

Bolli. United States Patent, 12; 2014.

Ballow M. Safety of IGIV therapy and infusion-related adverse events. Immunol Res. 2007;38(1-3):122-32. doi: 10.1007/s12026-007-0003-5, PMID 17917017

Chaudhary SB, Patel SB. Use of Amino Acids as Stabilizing Compounds in Pharmaceutical Compositions Containing High Concentrations of Protein-based Therapeutic Agents. US Patent no. US 2022/0087939A1; 24 March 2022.

Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949;177(2):751-66. doi: 10.1016/S0021-9258(18)57021-6, PMID 18110453

Aguirre J. The Kjeldahl Method: 140 Years. Switzerland: Springer Nature; 2023.

Indian Pharmacopeia Commission. Indian Pharmacopoeia. Vol. 1. Ch. 2.3.30, Ghaziabad: The Indian Pharmacopoeia Commission; 2022. p. 179-81.

Indian Pharmacopeia Commission. Monograph: Human normal immunoglobulin for intravenous use. In: Indian Pharmacopoeia. Vol. 3. Ghaziabad: The Indian Pharmacopoeia Commission; 2022. p. 4544-51.

Mariotti F, Tomeé D, Mirand PP. Converting nitrogen into protein-beyond 6.25 and Joness factors. Crit Rev Food Sci Nutr. 2008;48(2):177-84. doi: 10.1080/10408390701279749