EVALUATION OF EFFECTIVE PARAMETERS ON THE CONCENTRATING OF ANTI-RH (D) IMMUNOGLOBULIN PREPARATIONS USING ULTRAFILTRATION SYSTEM

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HASHEMI M.; JAFARI M.R.; BARANZADEH N.; VARASTEH A.R.

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Journal: Iranian Journal of Basic Medical Sciences Year:2007 | Volume:10 | Issue:1 (33) Page(s): 66-74

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Title

EVALUATION OF EFFECTIVE PARAMETERS ON THE CONCENTRATING OF ANTI-RH (D) IMMUNOGLOBULIN PREPARATIONS USING ULTRAFILTRATION SYSTEM

Author(s)

HASHEMI M. | JAFARI M.R. | BARANZADEH N. | VARASTEH A.R. | Issue Writer Certificate

Keywords

ANTI-RH (D)Q4

ULTRAFILTRATIONQ3

BIOACTIVITYQ2

ELATQ3

STABILITYQ2

IMMUNOGLOBULINQ3

Abstract

Anti-Rh (D) IMMUNOGLOBULIN is used for the prevention of Anti-Rh antibody production in Rh-individuals who have been exposed to Rh+ red blood cells. The purification and preparation processes of this product can directly effect its shelf life. Also, during the purification process using column chromatography methods, protein solutions are diluted and lose some of their STABILITY due to adsorption and fragmentation. Therefore, they should be concentrated with a proper procedure. The objective of this study was to concentrate the Anti-Rh (D) preparation using ULTRAFILTRATION system and optimize the conditions during concentrating process.Materials and Methods For ULTRAFILTRATION studies, 100ml of purified Anti-Rh(D) solutions containing 0.3 M Glycine and different concentrations of sucrose (30, 60, 90 mM) were concentrated to 27 ml in 50 min using Minitan-S system (filter type: polysulfon, cut off 30,000 Daltons) and pressure of 80 kPs. The clean water flux was used for the determination of suitable pump rate. Bradford protein assay was used for the quantification of total proteins in filtrate, SRID (Single Radial Immunodiffusion) for the determination of total IgG before and after concentrating of samples, gel filtration chromatography for the determination of the amount of aggregates and fragments and ELAT (Enzyme-Linked Antiglobulin Test) for the quantification of Anti- Rh (D) of the ultrafiltrated samples.ResultsWith mentioned conditions, the results of study showed that insignificant amount of protein and IgG transited from filter. Gel filtration studies using Sephadex G-200 showed that the amount of aggregates, fragments and monomer molecules in the concentrated Anti-Rh (D) solutions were 2.31%, 2.5% and 95%, respectively. The ELAT test showed that the amount of Anti-Rh (D) in concentrated-unformulated sample was reduced about 8%. For formulations containing 0.3 M Glycine, and sucrose with concentrations of 30, 60 and 90 mM, in phosphate buffer (pH 7.5,25 mM), the amounts of loss of Anti- Rh(D) were 2.3%,0.7% and 1.6%, respectively.Conclusion This study showed that ULTRAFILTRATION with the mentioned conditions is suitable for the concentrating of Anti-Rh (D) solutions. To prevent denaturation of IgG during ULTRAFILTRATION, it is better to add stabilizers, such as, Glycine and sucrose to Anti-Rh (D) IMMUNOGLOBULIN solutions before the process of ULTRAFILTRATION.

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APA: Copy

HASHEMI, M., JAFARI, M.R., BARANZADEH, N., & VARASTEH, A.R.. (2007). EVALUATION OF EFFECTIVE PARAMETERS ON THE CONCENTRATING OF ANTI-RH (D) IMMUNOGLOBULIN PREPARATIONS USING ULTRAFILTRATION SYSTEM. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 10(1 (33)), 66-74. SID. https://sid.ir/paper/52426/en

Vancouver: Copy

HASHEMI M., JAFARI M.R., BARANZADEH N., VARASTEH A.R.. EVALUATION OF EFFECTIVE PARAMETERS ON THE CONCENTRATING OF ANTI-RH (D) IMMUNOGLOBULIN PREPARATIONS USING ULTRAFILTRATION SYSTEM. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2007;10(1 (33)):66-74. Available from: https://sid.ir/paper/52426/en

IEEE: Copy

M. HASHEMI, M.R. JAFARI, N. BARANZADEH, and A.R. VARASTEH, “EVALUATION OF EFFECTIVE PARAMETERS ON THE CONCENTRATING OF ANTI-RH (D) IMMUNOGLOBULIN PREPARATIONS USING ULTRAFILTRATION SYSTEM,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 10, no. 1 (33), pp. 66–74, 2007, [Online]. Available: https://sid.ir/paper/52426/en

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