کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6136123 | 1224973 | 2008 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Novel human 3-domain disulfide-stabilized antibody fragment against glycoprotein of rabies virus
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کلمات کلیدی
PBSscFvRFFITCH1dsFvMTT - MTTRecombinant expression - بیان ترکیبیELISA - تست الیزاEnzyme-linked immunosorbent assay - تست الیزاStability - ثباتNeutralization - خنثی سازیsingle-chain variable fragment - قطعه متغیر تک زنجیره ایPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریvariable region of heavy chain - منطقه متغیر زنجیره سنگینRabies virus - ویروس هاری
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ایمونولوژی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Mutated disulfide bond sites VH (Cys44) and VL (Cys100) were constructed in variable domains (Fvs) of the human anti-glycoprotein antigen of the rabies virus (anti-GPRV), and the light chain variable (VL) and heavy chain variable (VH) fragments were linked using the heavy chain constant region 1 (CH1) of the human immunoglobulin (Ig) to successfully construct a 3-domain disulfide-stabilized fragment of variables (3d-dsFv). 3d-dsFv was mainly expressed as an inclusion body. After refolding by the conventional dilution method, 3d-dsFv was purified using a nickel-nitrilotriacetic acid (Ni-NTA) column. Enyzme-linked immunosorbent assay (ELISA) was used to determine the binding activity of 3d-dsFv to GPRV. Flow cytometry studies and rapid fluorescent focus inhibition test were used to evaluate the function of 3d-dsFv. The results showed that the stability of 3d-dsFv was improved notably in some aspects such as thermal kinetics, ability to withstand urea denaturation, etc. 3d-dsFv could bind specially to infective cells and the GPRV. The titration of 3d-dsFv to RV-CVS is 83.3 IU/mg, and it can easily reach 2.5Â IU/mL, which is the value suggested by the WHO as effective for neutralization titration of the rabies virus.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Microbes and Infection - Volume 10, Issue 5, April 2008, Pages 548-555
Journal: Microbes and Infection - Volume 10, Issue 5, April 2008, Pages 548-555
نویسندگان
Kun Cai, Hui Wang, Shizhong Bao, Jing Shi, Xiaojun Hou, Xiang Gao, Hao Liu, Jun Yin,