Isoenzyme-directed selection and characterization of anti-creatine kinase single chain Fv antibodies from a human phage display library

2002 ◽  
Vol 1579 (2-3) ◽  
pp. 124-132 ◽  
Author(s):  
Uwe Schlattner ◽  
Christof Reinhart ◽  
Thorsten Hornemann ◽  
Malgorzata Tokarska-Schlattner ◽  
Theo Wallimann
Keyword(s):  
Creatine Kinase ◽  
Phage Display ◽  
Phage Display Library ◽  
Single Chain ◽  
Single Chain Fv

scholarly journals Characterization of LL25X, a Newly Isolated Anti-SIV Monoclonal Antibody, to Determine Binding Specificity

Elements ◽  
2017 ◽  
Vol 13 (1) ◽  
Author(s):  
Zackary Tajin Park
Keyword(s):  
Phage Display ◽  
Mammalian Cells ◽  
Expression Vector ◽  
Restriction Enzymes ◽  
Phage Display Library ◽  
Single Chain ◽  
Phage Display Technology ◽  
Mammalian Expression ◽  
Mammalian Expression Vector ◽  
Single Chain Fv

A phage display library was previously constructed from an SIV-infected rhesus macaque. Several single chain Fv (scFv), including SU24, SU343 and LL25X, were selected using phage display technology. Sequences corresponding to SU24, SU343 and LL25X were optimized for expression in a mammalian system and commercially synthesized. SU24 and SU343 had previously been cloned into a mammalian expression vector. In this study, we aimed to characterize the specificity of SU24, SU343, and LL25X.. The codon-optimized version of the scFv LL25X gene sequence was cloned into a mammalian expression vector (pCEP4).  LL25X DNA was amplified by PCR, and the PCR product and mammalian expression vector were both digested with KpnI/SapI restriction enzymes. Digested fragments were purified, and the fragments were ligated using T4DNA ligase. E. coli cells were transformed with the ligation reaction. Single colonies were selected on LB agar plates containing the selective antibiotic (ampicillin). Positive colonies were identified after DNA mini-preparation and test-digestion with KpnI and SapI. Sanger sequencing confirmed cloning results and DNA sequence accuracy. Following transfection of mammalian cells (293T), LL25X-Fc cells, and purifying our protein, the binding of LL25X-Fc to the SIV gp140 envelope protein was confirmed via ELISA and Western Blotting.

Characterization of anti-mouse FcγRII single-chain Fv fragments derived from human phage display libraries

1998 ◽  
Vol 4 (1) ◽  
pp. 71-87 ◽  
Author(s):  
A.M McCall ◽  
A.R Amoroso ◽  
C Sautès ◽  
J.D Marks ◽  
L.M Weiner
Keyword(s):  
Phage Display ◽  
Single Chain ◽  
Single Chain Fv ◽  
Phage Display Libraries

scholarly journals Screening for a human single chain Fv antibody against epitope on amyloid-beta 1-40 from a human phage display library

2007 ◽  
Vol 120 (21) ◽  
pp. 1931-1934 ◽  
Author(s):  
Zhen-fu ZHAO ◽  
Guo-quan GAO ◽  
Shu LIU ◽  
Jun-tao ZOU ◽  
Yao XIE ◽  
...  
Keyword(s):  
Phage Display ◽  
Amyloid Beta ◽  
Phage Display Library ◽  
Single Chain ◽  
Single Chain Fv ◽  
Fv Antibody ◽  
Single Chain Fv Antibody
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