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Faculty of Science and Arts - Khulais
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 Type
Dr Grade
Email
عمرو السيد الحكيم
El Hakim, Amr El Sayed
Researcher
Doctorate
رجاء رضا حامد
Hamed, Ragaa Reda
Researcher
Doctorate
طارق محمد مصطفى
Mostafa, Tarek Mohamed
Researcher
Doctorate
عبد العزيز محمد جاد
Gad, Abdel Aziz mohamed
Researcher
Master
Files
File Name
Type
Description
28630.pdf
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