EFFECT OF AMINO ACID ADDITION AS STABILIZERS ON TOTAL PROTEIN CONTENT OF INTRAVENOUS THERAPEUTIC HUMAN NORMAL IMMUNOGLOBULIN
EFFECT OF AMINO ACID STABILIZERS ON PROTEIN CONTENT OF THERAPEUTIC HUMAN NORMAL IMMUNOGLOBULIN (IV)
Keywords:
Human immunoglobulin, Quality control, Protein estimation, Kjeldahl, BiuretAbstract
Objectives: This study was conducted to show the variation in total protein content of therapeutic immunoglobulin batches after 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 method. The total protein in these samples was also determined by both methods after addition of 0.3 M Glycine, 0.3 M Arginine, 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 or Arginine or Histidine has been added. The total protein content by Kjeldahl method in the samples added with glycine, or arginine or histidine was found higher as compared to Biuret method.
Conclusion: Our study suggests that the manufacturers of therapeutic immunoglobulin should replace amino acid stabilizer with a non-nitrogenous stabilizer, because the Kjeldahl method recommended in various pharmacopoeias is based on detection of total nitrogen. Therefore, 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 pharmacopoeia.
Downloads
References
Stiehm ER. Human gamma globulins as therapeutic agents. Adv Pediatr 1988;35:1-72.
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. 1, London, 1999. Core SPC for human normal immunoglobulin for subcutaneous and intramuscular use (SC/IMIg) CPMP/ BPWG/282/00, London;2002.
Pharmacopoea Europaea, 3. (Europaisches Arzneibuch) Amtliche Deutsche Ausgabe. DAV, Stuttgart, p 1289;Ausgabe 1997
Gaines A, Varricchio F, Kapit R. Renal insufficiency and failure associated with immune globulin intravenous therapy–United States, 1985–1998. MMWR 1999;48: 518-21.
Patil V, Kaveri SV. The mechanisms of action of IVIG in autoimmune and inflammatory diseases. ISBT Sci Ser 2013;8:185-8.
Szenczi A, Kardos J, Medgyesi GA, Zavodszky P. The effect of solvent environment on the conformation and stability of human polyclonal IgG in solution. Biologicals 2006;34:5-14.
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 2012;173:1-19.
Bolli. United States Patent, 12, 2014.
Ballow M. Safety of IGIV therapy and infusion-related adverse events. Immunol Res 2007;38:122-32.
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 2022/0087939 A1, 24 March 2022.
Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem 1949; 177:751-66.
Aguirre J. The Kjeldahl Method: 140 years, Springer Nature, Switzerland, 2023.
Indian Pharmacopeia Commission, Indian Pharmacopoeia, Vol. 1, Chapter 2.3.30, Method F, Page No. 179-81, 2022.
Indian Pharmacopeia Commission, Indian Pharmacopoeia, Vol. III, Monograph: Human normal immunoglobulin for intravenous use, Page No. 4544-51, 2022.
Mariotti F, Tome D, Mirand PP. Converting nitrogen into protein-beyond 6.25 and Joness factors. Crit Rev Food Sci Nutr 2008;48:177-84.
Published
How to Cite
Issue
Section
Copyright (c) 2024 Dr. MANOJ Kumar Rajput, Manvandra Pratap Singh, Aprajita Singh, Tara Chand, Anoop Kumar, Meena Kumari
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.