Selection and characterization of single chain Fv fragments against murine recombinant prion protein from a synthetic human antibody phage display library

AbstractSARS-CoV-2 (Covid-19) has caused currently ongoing global plague and imposed great challenges to health managing systems all over the world, with millions of infections and hundreds of thousands of deaths. In addition to racing to develop vaccines, neutralizing antibodies (nAbs) to this virus have been extensively sought and are expected to provide another prevention and therapy tool against this frantic pandemic. To offer fast isolation and shortened early development, a large human naïve phage display antibody library, was built and used to screen specific nAbs to the receptor-binding domain, RBD, the key for Covid-19 virus entry through a human receptor, ACE2. The obtained RBD-specific antibodies were characterized by epitope mapping, FACS and neutralization assay. Some of the antibodies demonstrated spike-neutralizing property and ACE2-competitiveness. Our work proved that RBD-specific neutralizing binders from human naïve antibody phage display library are promising candidates to for further Covid-19 therapeutics development.

Download Full-text

Neutralizing Antibodies of Botulinum Neurotoxin Serotype A Screened from a Fully Synthetic Human Antibody Phage Display Library

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057109343206 ◽

2009 ◽

Vol 14 (8) ◽

pp. 991-998 ◽

Cited By ~ 18

Author(s):

Rui Yu ◽

Shuang Wang ◽

Yun-zhou Yu ◽

Wei-shi Du ◽

Fang Yang ◽

...

Keyword(s):

Phage Display ◽

Neutralizing Antibodies ◽

Botulinum Neurotoxin ◽

Phage Display Library ◽

Human Antibody ◽

Serotype A ◽

Antibody Phage ◽

Antibody Phage Display ◽

Botulinum Neurotoxin Serotype A

The botulinum neurotoxins (BoNTs) produced by Clostridium botulinum are the most poisonous protein substances known. The neutralizing antibodies against botulinum neurotoxin can effectively prevent and cure the toxicosis. Using purified Hc fragments of botulinum neurotoxin serotype A (BoNT/A-Hc) as antigen, 2 specific neutralizing antibodies mapping different epitopes were selected from a fully synthetic human antibody library. The 2 antibodies can effectively inhibit the binding between BoNT/A-Hc and differentiated PC-12 cells in vitro, and the neutralization was evaluated in vivo. Although no single mAb completely protected mice from toxin, they both could prolong time to death when challenged with 20 LD 50s (50% lethal doses) of BoNT/A. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, suggesting their high neutralizing potency in vivo . The results would lead to further production of neutralizing antibody drugs against BoNT/A. It also proved that it was a quick method to obtain human therapeutic antibodies by selecting from the fully synthetic human antibody phage display library. ( Journal of Biomolecular Screening 2009:991-998)

Download Full-text

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

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression ◽

10.1016/s0167-4781(02)00530-4 ◽

2002 ◽

Vol 1579 (2-3) ◽

pp. 124-132 ◽

Cited By ~ 12

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

Download Full-text

Specificity grafting of human antibody frameworks selected from a phage display library: generation of a highly stable humanized anti-CD22 single-chain Fv fragment

Protein Engineering Design and Selection ◽

10.1093/protein/gzg096 ◽

2003 ◽

Vol 16 (10) ◽

pp. 753-759 ◽

Cited By ~ 28

Author(s):

J. Krauss ◽

M. A.E. Arndt ◽

A. C.R. Martin ◽

H. Liu ◽

S. M. Rybak

Keyword(s):

Phage Display ◽

Phage Display Library ◽

Human Antibody ◽

Single Chain ◽

Single Chain Fv ◽

Single Chain Fv Fragment

Download Full-text

Screening of a human antibody phage display library against a peptide antigen using stimuli-responsive bioconjugates

Biotechnology Progress ◽

10.1002/btpr.60 ◽

2008 ◽

Vol 24 (6) ◽

pp. 1314-1324 ◽

Cited By ~ 2

Author(s):

Gisela Stocker ◽

Dorothée Dumoulin ◽

Caroline Vandevyver ◽

Frank Hilbrig ◽

Ruth Freitag

Keyword(s):

Phage Display ◽

Phage Display Library ◽

Human Antibody ◽

Stimuli Responsive ◽

Peptide Antigen ◽

Antibody Phage ◽

Antibody Phage Display

Download Full-text

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

Elements ◽

10.6017/eurj.v13i1.9604 ◽

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.

Download Full-text