Document Details

Document Type : Article In Journal 
Document Title :
Biochemical studies on the conjugation of antibodies with turnip peroxidase using two different methods
دراسات كيميائية حيوية على ارتباط الأجسام المناعية بانزيم البيروكسيدييز باستخدام طريقتين مختلفتين
 
Subject : Applied Science 
Document Language : English 
Abstract : The efficiency of conjugation between major carbohydrate rich peroxidase (TPOD) of turnip roots (Brassica rapa) and antibodies of rabbit anti-mouse IgGs (AM IgGs) was carried out for production of TPOD-AM IgG conjugate suitable for immunodiagnostic kits. The conjugate was prepared by two methods: one-step glutaraldehyde conjugation and periodate conjugation. The conjugates were characterized immunochemically and immuno-enzymatically. The two methods were comparable regarding the immunological activity of the conjugated products, while the ELISA titer of the conjugate prepared by the periodate method was almost double that prepared by the glutaraldehyde method. The conjugated product prepared by the one-step glutaraldehyde method was stable for at least six months at –20ÆC with less than 20% loss of activity. Characterization of the TPOD and TPOD-AM IgG conjugate including Km values, optimum pH, optimum temperature, and thermal stability were carried out. The Km values of the TPOD and TPOD-AM IgG conjugate (ABTS) were 1.2 and 0.86 mM at pH 4 and 0.52 and 0.6 mM at pH 6, respectively. The Km values of these two products (H2O2) were 2 and 1.25 mM at pH 4 and 0.35 and 1.2 mM at pH 6, respectively. The optimum pH value was 3.5 for both while optimum temperature values were 55 and 50ÆC, respectively. Economically, the conjugated product by the one-step glutaraldehyde method was preferable, as it requires one step for its purification with higher recovery, and immunologically comparable beside its higher stability compared with the conjugate prepared by the periodate method. 
ISSN : 1790-045X 
Journal Name : Journal of Biological Research-Thessaloniki 
Volume : 12 
Issue Number : 2 
Publishing Year : 1430 AH
2009 AD 
Article Type : Article 
Added Date : Saturday, January 15, 2011 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
عمرو السيد الحكيمEl Hakim, Amr El SayedResearcherDoctorate 
رجاء رضا حامدHamed, Ragaa RedaResearcherDoctorate 
طارق محمد مصطفىMostafa, Tarek MohamedResearcherDoctorate 
عبد العزيز محمد جادGad, Abdel Aziz mohamedResearcherMaster 

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