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:: Volume 19, Issue 2 (Summer 2022) ::
Sci J Iran Blood Transfus Organ 2022, 19(2): 108-121 Back to browse issues page
Evaluation of effective factors during an in Vitro immunization against Kell blood group antigen
F. Tobeyani , S. Milani , F. Yari *
Keywords: Key words: Immunization, Antibody, Kell Blood-group System
Full-Text [PDF 745 kb]   (692 Downloads)     |   Abstract (HTML)  (1135 Views)
Type of Study: Research | Subject: Imunohematology
Published: 2022/07/1
Full-Text:   (715 Views)
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Sci J Iran Blood Transfus Organ 2022;19 (2): 108-121
Original Article
 



Evaluation of effective factors during an
in Vitro immunization against Kell blood group antigen

Tobeyani F.1, Milani S.1, Yari F.1

1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran



Abstract
Background and Objectives
in-vitro immunization is one of the common methods of immunization as a preliminary step in the production of Monoclonal Antibodies. In this method, peripheral blood mononuclear cells (PBMC) are isolated and incubated in culture medium with necessary factors for immunization. Antigen-specific B lymphocytes proliferate and differentiate into cells that secrete specific antibodies. In the current study, we aimed to identify various factors and contributing elements on B cell In-vitro immunization process for the generation of antibody against Kell blood group antigen.

Materials and Methods
In this experimental study, Kell antigen was purified from Kell+ whole blood. Peripheral blood mononuclear cells were isolated using a density gradient approach from Kell- whole blood bag. After exposure to L-Leucyl-L-leucine methyl ester (LLME), they were incubated with other required factors for immunization, including antigen (500-1000 ng/ml), adjuvant (5-20 mg/ml), and cytokines (9.5 pg/ml). After 7-11 days, cells supernatant was evaluated using a sensitive method, ELISA, for antibody production.

Results
It was observed that the most desirable culture condition for immunization and further production of specific anti-Kell antibody from mononuclear cells was 0.25 mM concentration of LLME employing 500-1000 ng of Kell antigens and 10 microliter MLC. In the mentioned condition, the optical density was read at 450 nm as 2.052 ± 0.03, which was the representative of production of specific anti-Kell antibody.

Conclusions 
Based on the study, immunization in culture medium against Kell antigen can be done. By employing different factors, the production of anti-Kell producing cells in culture condition can be optimized.

Key words: Immunization, Antibody, Kell Blood-group System




Received:  17 Jan  2022
Accepted: 13 Mar 2022



Correspondence: Yari F., PhD of Immunology. Professor of Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine.
P.O.Box: 14665-1157, Tehran, Iran. Tel: (+9821) 82052237; Fax: (+9821) 88601555
E-mail: f.yari@ibto.ir
 
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Tobeyani F, Milani S, Yari F. Evaluation of effective factors during an in Vitro immunization against Kell blood group antigen. Sci J Iran Blood Transfus Organ 2022; 19 (2) :108-121
URL: http://bloodjournal.ir/article-1-1441-en.html
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Volume 19, Issue 2 (Summer 2022) Back to browse issues page
فصلنامه پژوهشی خون Scientific Journal of Iran Blood Transfus Organ
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