Generation and Characterization of Anti-chitinase Monoclonal Antibodies

被引：9
|
作者：
Soukhtanloo, Mohammad [2 ]
Falak, Reza [1 ]
Sankian, Mojtaba [1 ]
Varasteh, A-Reza [3 ]
机构：
[1] Mashhad Univ Med Sci, Immunol Res Ctr, Bu Ali Res Inst, Mashhad 96911, Khorasan Razavi, Iran
[2] Mashhad Univ Med Sci, Fac Med, Dept Biochem, Mashhad 96911, Khorasan Razavi, Iran
[3] Mashhad Univ Med Sci, Varastegan Inst Med Sci, Immunobiochem Lab, Mashhad 96911, Khorasan Razavi, Iran
来源：
HYBRIDOMA | 2011年 / 30卷 / 02期
关键词：
LIPID-TRANSFER PROTEIN; DUST MITE ALLERGEN; MAJOR ALLERGEN; MOLECULAR-CLONING; IGE-BINDING; IN-VIVO; POLLEN; GRAPE; STANDARDIZATION; IDENTIFICATION;
D O I：
10.1089/hyb.2010.0088
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Class IV chitinase, an allergenic protein of Vitis vinifera (grape), was purified by anion exchange chromatography and used for immunization of Balb/c mice. Monoclonal antibodies (MAbs) were raised by hybridoma technology using Sp2/0 myeloma cells. Finally after three limiting dilutions, six stable clones were generated. Antibody isotyping showed that IgG(2a), IgG(2b), and IgM were produced by one, two, and three of the clones, respectively. All of the MAbs had kappa light chain. The affinities were in the range of 3 x 10(8) to 1.2 x 10(9) M-1. The MAbs were specific for grape chitinase as confirmed by Western blotting. In conclusion, we successfully produced several MAbs against grape class IV chitinase, which could be used for assessment of this allergen in different grape cultivars.
引用
收藏
页码：145 / 151
页数：7
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