Protective anti-prion antibodies in human immunoglobulin repertoires

AbstractPrion immunotherapy may hold great potential, but antibodies against certain PrP epitopes can be neurotoxic. Here we identified >6000 PrP-binding antibodies in a synthetic human Fab phage display library, 49 of which we characterized in detail. Antibodies directed against the flexible tail of PrP conferred neuroprotection against infectious prions. We then mined published repertoires of circulating B cells from healthy humans and found antibodies similar to the protective phage-derived antibodies. When expressed recombinantly, these antibodies exhibited anti-PrP reactivity. Furthermore, we surveyed 48’718 samples from 37’894 hospital patients for the presence of anti-PrP IgGs, and found 21 high-titer individuals. The clinical files of these individuals did not reveal any enrichment of specific pathologies, suggesting that anti-PrP autoimmunity is innocuous. The existence of protective anti-prion antibodies in unbiased human immunological repertoires, combined with the reported lack of such antibodies in carriers of disease-associated PRNP mutations, suggests a link to the low incidence of spontaneous prion diseases in human populations.

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Use of cyclic peptide phage display library for the identification of a CD45RC epitope expressed on murine B cells and their precursors

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(03)01264-6 ◽

2003 ◽

Vol 307 (4) ◽

pp. 791-796 ◽

Cited By ~ 6

Author(s):

Tamás Czömpöly ◽

Árpád Lábadi ◽

Mercedesz Balázs ◽

Péter Németh ◽

Péter Balogh

Keyword(s):

B Cells ◽

Phage Display ◽

Cyclic Peptide ◽

Phage Display Library ◽

Peptide Phage Display

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Detection of Cystic Fibrosis Serological Biomarkers Using a T7 Phage Display Library

Scientific Reports ◽

10.1038/s41598-017-18041-2 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 3

Author(s):

Harvinder Talwar ◽

Samer Najeeb Hanoudi ◽

Andreea Geamanu ◽

Dana Kissner ◽

Sorin Draghici ◽

...

Keyword(s):

Cystic Fibrosis ◽

Phage Display ◽

Phage Display Library ◽

T7 Phage Display ◽

Serological Biomarkers ◽

T7 Phage

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Production, characterization and in-vitro applications of single-domain antibody against thyroglobulin selected from novel T7 phage display library

Journal of Immunological Methods ◽

10.1016/j.jim.2021.112990 ◽

2021 ◽

Vol 492 ◽

pp. 112990

Author(s):

Jothivel Kumarasamy ◽

Samar Kumar Ghorui ◽

Chandrakala Gholve ◽

Bharti Jain ◽

Yogesh Dhekale ◽

...

Keyword(s):

Phage Display ◽

Phage Display Library ◽

Single Domain ◽

Single Domain Antibody ◽

Domain Antibody ◽

T7 Phage Display ◽

T7 Phage

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Stable heterodimers from remodeling the domain interface of a homodimer using a phage display library

Journal of Molecular Biology ◽

10.1006/jmbi.1997.1116 ◽

1997 ◽

Vol 270 (1) ◽

pp. 26-35 ◽

Cited By ~ 145

Author(s):

Shane Atwell ◽

John B.B Ridgway ◽

James A Wells ◽

Paul Carter

Keyword(s):

Phage Display ◽

Phage Display Library

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Isolation of a highly specific ligand for the alpha 5 beta 1 integrin from a phage display library

The Journal of Cell Biology ◽

10.1083/jcb.124.3.373 ◽

1994 ◽

Vol 124 (3) ◽

pp. 373-380 ◽

Cited By ~ 225

Author(s):

E Koivunen ◽

B Wang ◽

E Ruoslahti

Keyword(s):

Phage Display ◽

Cyclic Peptide ◽

Cell Attachment ◽

Phage Display Library ◽

Cell Binding ◽

Rgd Peptides ◽

Beta 1 Integrin ◽

Alpha V Beta 3 ◽

Integrin Ligand ◽

Specific Ligand

Our previous studies showed that the alpha 5 beta 1 integrin selects cysteine pair-containing RGD peptides from a phage display library based on a random hexapeptide. We have therefore searched for more selective peptides for this integrin using a larger phage display library, where heptapeptides are flanked by cysteine residues, thus making the inserts potentially cyclic. Most of the phage sequences that bound to alpha 5 beta 1 (69 of 125) contained the RGD motif. Some of the heptapeptides contained an NGR motif. As the NGR sequence occurs in the cell-binding region of the fibronectin molecule, this sequence could contribute to the specific recognition of fibronectin by alpha 5 beta 1. Selection for high affinity peptides for alpha 5 beta 1 surprisingly yielded a sequence RRETAWA that does not bear obvious resemblance to known integrin ligand sequences. The synthetic cyclic peptide GACRRETAWACGA (*CRRETAWAC*) was a potent inhibitor of alpha 5 beta 1-mediated cell attachment to fibronectin. This peptide is nearly specific for the alpha 5 beta 1 integrin, because much higher concentrations were needed to inhibit the alpha v beta 1 integrin, and there was no effect on alpha v beta 3- and alpha v beta 5-mediated cell attachment to vitronectin. The peptide also did not bind to the alpha IIb beta 3 integrin. *CRRETAWAC* appears to interact with the same or an overlapping binding site in alpha 5 beta 1 as RGD, because cell attachment to *CRRETAWAC* coated on plastic was divalent cation dependent and could be blocked by an RGD-containing peptide. These results reveal a novel binding specificity in the alpha 5 beta 1 integrin.

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Molecular characteristics of antibodies bearing an anti-DNA-associated idiotype.

Journal of Experimental Medicine ◽

10.1084/jem.174.6.1639 ◽

1991 ◽

Vol 174 (6) ◽

pp. 1639-1652 ◽

Cited By ~ 53

Author(s):

A Manheimer-Lory ◽

J B Katz ◽

M Pillinger ◽

C Ghossein ◽

A Smith ◽

...

Keyword(s):

B Cells ◽

Dna Binding ◽

Lupus Erythematosus ◽

Germ Line ◽

High Titer ◽

Variable Region ◽

Amino Acid Sequences ◽

Molecular Characteristics ◽

Systemic Lupus ◽

Dna Antibodies

Anti-double-stranded DNA antibodies are the hallmark of the disease systemic lupus erythematosus and are believed to contribute to pathogenesis. While a large number of anti-DNA antibodies from mice with lupus-like syndromes have been characterized and their variable region genes sequenced, few human anti-DNA antibodies have been reported. We describe here the variable region gene sequences of eight antibodies produced by Epstein-Barr virus (EBV)-transformed B cells that bear the 3I idiotype, an idiotype expressed on anti-DNA antibodies and present in high titer in patients with systemic lupus. The comparison of these antibodies to the light chains of 3I+ myeloma proteins and serum antibodies reveals that EBV transformation yields B cells producing antibodies representative of the expressed antibody repertoire. The analysis of nucleotide and amino acid sequences of these antibodies suggests the first complementarity determining region of the light chain may be important in DNA binding and that paradigms previously generated to account for DNA binding require modification. The understanding of the molecular genetics of the anti-DNA response requires a more complete description of the immunoglobulin germ line repertoire, but data reported here suggest that somatic diversification is a characteristic of the anti-DNA response.

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Antibody design using LSTM based deep generative model from phage display library for affinity maturation

Scientific Reports ◽

10.1038/s41598-021-85274-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Koichiro Saka ◽

Taro Kakuzaki ◽

Shoichi Metsugi ◽

Daiki Kashiwagi ◽

Kenji Yoshida ◽

...

Keyword(s):

Phage Display ◽

Short Term Memory ◽

Current Method ◽

Phage Display Library ◽

Affinity Maturation ◽

Generative Model ◽

Sequence Generation ◽

Machine Learning Approach ◽

Antibody Design ◽

Ngs Data

AbstractMolecular evolution is an important step in the development of therapeutic antibodies. However, the current method of affinity maturation is overly costly and labor-intensive because of the repetitive mutation experiments needed to adequately explore sequence space. Here, we employed a long short term memory network (LSTM)—a widely used deep generative model—based sequence generation and prioritization procedure to efficiently discover antibody sequences with higher affinity. We applied our method to the affinity maturation of antibodies against kynurenine, which is a metabolite related to the niacin synthesis pathway. Kynurenine binding sequences were enriched through phage display panning using a kynurenine-binding oriented human synthetic Fab library. We defined binding antibodies using a sequence repertoire from the NGS data to train the LSTM model. We confirmed that likelihood of generated sequences from a trained LSTM correlated well with binding affinity. The affinity of generated sequences are over 1800-fold higher than that of the parental clone. Moreover, compared to frequency based screening using the same dataset, our machine learning approach generated sequences with greater affinity.

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