A Fibrin-Specific Monoclonal Antibody from a Designed Phage Display Library Inhibits Clot Formation and Localizes to Tumors In Vivo

2014 ◽  
Vol 426 (21) ◽  
pp. 3606-3618 ◽  
Author(s):  
Alessia Putelli ◽  
Jonathan D. Kiefer ◽  
Matthias Zadory ◽  
Mattia Matasci ◽  
Dario Neri
Keyword(s):  
Monoclonal Antibody ◽  
Phage Display ◽  
Phage Display Library ◽  
Clot Formation ◽  
Specific Monoclonal Antibody

scholarly journals A novel and effective approach to generate germline-like monoclonal antibodies by integration of phage and mammalian cell display platforms

2021 ◽  
Author(s):  
Yu-jia Jin ◽  
Diao Yu ◽  
Xiao-long Tian ◽  
Hui-xian Li ◽  
Xiao-chao Zhou ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Phage Display ◽  
Mammalian Cell ◽  
Phage Display Library ◽  
Antibody Fragments ◽  
Human Antibody ◽  
Phage Display Technology ◽  
Mammalian Cell Display

AbstractPhage display technology allows for rapid selection of antibodies from the large repertoire of human antibody fragments displayed on phages. However, antibody fragments should be converted to IgG for biological characterizations and affinity of antibodies obtained from phage display library is frequently not sufficient for efficient use in clinical settings. Here, we describe a new approach that combines phage and mammalian cell display, enabling simultaneous affinity screening of full-length IgG antibodies. Using this strategy, we successfully obtained a novel germline-like anti-TIM-3 monoclonal antibody named m101, which was revealed to be a potent anti-TIM-3 therapeutic monoclonal antibody via in vitro and in vivo experiments, indicating its effectiveness and power. Thus, this platform can help develop new monoclonal antibody therapeutics with high affinity and low immunogenicity.

scholarly journals Efficient In Vivo Selection of a Novel Tumor-Associated Peptide from a Phage Display Library

Molecules ◽  
2011 ◽  
Vol 16 (1) ◽  
pp. 900-914 ◽  
Author(s):  
Anka N. Veleva ◽  
Desh B. Nepal ◽  
C. Brandon Frederick ◽  
Jacob Schwab ◽  
Pamela Lockyer ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Library ◽  
In Vivo Selection ◽  
Selection Of

Abstract 12105: Targeted Delivery of Therapeutic Agents After Myocardial Infarction

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Siva Sai Krishna Dasa ◽  
Marc E Seamen ◽  
Brent A French ◽  
Kimberly A Kelly
Keyword(s):  
Myocardial Infarction ◽  
Phage Display ◽  
Targeted Delivery ◽  
Cell Types ◽  
Phage Display Library ◽  
Border Zone ◽  
Cell Type ◽  
Cellular Targets ◽  
Cell Type Specific

Introduction: Current therapies for heart failure (HF) after myocardial infarction (MI) only slow the progression of LV remodeling and have little capacity to regenerate cardiac muscle lost to MI. To expedite targeted delivery of regenerative therapies post-MI, we hypothesized that suitable targets could be identified by biopanning the heart with a phage display library in a mouse model of MI. Methods: A phage display library was biopanned in vivo to identify peptides specific for the infarct/border zone 4 days post-MI. Fluorescence molecular tomography (FMT) followed by tissue immunofluorescence was performed to interrogate the specificity of phage groups and individual clones with targeted phage at VT680 and neg control phage at VT750. The VT680 fluorophore on the targeted phage clones was then used to identify the cellular targets of those clones by counter-staining with antibodies against cell types of interest. Results: We identified phage clones specific for endothelium, cardiomyocytes, inflammatory fibroblasts and c-Kit+ cells present in the border zone post-MI. Liposomes conjugated with different cell type specific peptides had different accumulation rates in the post-infarct heart as visualized by FMT imaging (Fig. 1a). Immunofluorescence analysis demonstrated cell-type specific association of the targeted liposomes with cells expressing c-Kit, CD31 and Hrnr (Figs. 1b&c). We have also been successful in remote loading of anti-apoptotic and immune suppresive drugs into these liposomes and are currently studying their effect in mice after MI. Conclusions: Peptides identified by this screen enable the targeting of different cell types present in the border zone with different drugs. Identifying the molecular binding partners for these peptides may yield insight into the various events/pathways that evolve after a myocardial infarction.

scholarly journals Development of a cleavage-site-specific monoclonal antibody for detecting metalloproteinase-derived aggrecan fragments: detection of fragments in human synovial fluids

1995 ◽  
Vol 310 (1) ◽  
pp. 337-343 ◽  
Author(s):  
A J Fosang ◽  
K Last ◽  
P Gardiner ◽  
D C Jackson ◽  
L Brown
Keyword(s):  
Monoclonal Antibody ◽  
Broad Band ◽  
Specific Monoclonal Antibody ◽  
Site Specific ◽  
Molar Basis ◽  
Dominant Band ◽  
Synovial Fluids ◽  
Neutrophil Collagenase ◽  
Phenylalanine Residue

We have developed a monoclonal antibody AF-28 that specifically recognizes a neo-epitope on polypeptides with N-terminal FFGVG ... sequences. This sequence is found at the N-terminus of aggrecan fragments that have been digested with matrix metalloproteinases (MMPs). By immunoblotting, monoclonal antibody AF-28 specifically detected G2 fragments derived from an aggrecan G1-G2 substrate digested with stromelysin, collagenase, gelatinase and matrilysin, but failed to detect G2 fragments obtained from elastase, trypsin or cathepsin B digests. Undigested G1-G2 was not detected. In addition, AF-28 antibody detected fragments derived from whole aggrecan and this detection did not require prior treatment with chondroitinase or keratanase. Competition experiments confirmed that peptides containing internal ... FFGVG ... sequences were not detected by the antibody, while native MMP-digested aggrecan fragments and a synthetic 32-mer peptide with FFGVG ... N-termini were equally competitive on a molar basis. An FFGVG 5-mer, and an FGVGGEEDI9-mer which lacked the N-terminal phenylalanine residue, were 50 times and 230 times respectively less competitive than the FFGVG ... 32-mer. Two fragments from the interglobular domain, F342-F373 and F342-D441, that are predicted products of G1-G2 digestion by neutrophil collagenase but have not previously been detected, could be detected with AF-28. The epitope recognized by AF-28 was also detected in human synovial fluids by Western blot analysis. A broad band of 100-200 kDa was detected in some patients and a dominant band of 40-60 kDa was found in two patients. The size of this small fragment corresponds with that seen for the porcine F342-E373 product and may represent the natural physiological product of aggrecan cleaved in vivo at both the MMP site (... DIPEN341 decreases F342FGVG ...) and the aggrecanase site (... ITEGE373 decreases A374RGSVI ...).

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

2009 ◽  
Vol 14 (8) ◽  
pp. 991-998 ◽  
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)

Sign in / Sign up

Export Citation Format

Share Document