scholarly journals Generation of a Large Peptide Phage Display Library by Self-Ligation of Whole-Plasmid PCR Product

2020 ◽  
Vol 15 (11) ◽  
pp. 2907-2915
Author(s):  
Xu-Dong Kong ◽  
Vanessa Carle ◽  
Cristina Díaz-Perlas ◽  
Kaycie Butler ◽  
Christian Heinis
Keyword(s):  
Phage Display ◽  
Phage Display Library ◽  
Pcr Product ◽  
Peptide Phage Display ◽  
Large Peptide

scholarly journals Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: Specificity study using a peptide phage display library

2011 ◽  
Vol 181 (2-4) ◽  
pp. 291-300 ◽  
Author(s):  
Renan O. Clara ◽  
Tatiane S. Soares ◽  
Ricardo J.S. Torquato ◽  
Cássia A. Lima ◽  
Renata O.M. Watanabe ◽  
...  
Keyword(s):  
Phage Display ◽  
Cysteine Protease ◽  
Cathepsin L ◽  
Phage Display Library ◽  
Boophilus Microplus ◽  
Peptide Phage Display ◽  
Specificity Study ◽  
Protease Specificity

Screening and identification of the mimic epitope of the adhesion protein of Mycoplasma genitalium

2012 ◽  
Vol 58 (7) ◽  
pp. 898-908 ◽  
Author(s):  
Yanhua Zeng ◽  
Liangzhuan Liu ◽  
Jun He ◽  
Yan Liu ◽  
Cuiming Zhu ◽  
...  
Keyword(s):  
Amino Acid ◽  
Phage Display ◽  
Mycoplasma Genitalium ◽  
Phage Display Library ◽  
Antigenic Structure ◽  
Amino Acid Residues ◽  
Adhesion Protein ◽  
Phage Display Peptide Library ◽  
Screening And Identification ◽  
Peptide Phage Display

Mycoplasma genitalium adhesion protein (MgPa) is the major adhesion protein of M. genitalium, and its C-terminal domain (amino acid 1075–1444) is the most immunogenic region. However, the exact epitopes of the adhesion protein of M. genitalium are still unclear. We used the purified polyclonal antibody against the recombinant adhesion protein to screen the mimic epitopes of MgPa using a random 12-peptide phage display library. Immunoscreening via the phage display peptide library revealed that 3 motifs (P-S-A-A/V-X-R-F/W-E/S-L-S-P, A-K-I/L-T/Q-X-T-L-X-L, and K-S-L-S-R-X-D-X-I) may represent 3 different mimotopes of MgPa. Results of bioinformatics analysis by MIMOX demonstrated that the key consensus amino acid residues in the aligned mimotopes may be S, A, and F for cluster 1; A, K, I, T, and L for cluster 2; and K, S, L, R, D, and I for cluster 3. Three representative phages could recognize sera from M. genitalium-positive patients to varying degrees, whereas they could not recognize the sera from Mycoplasma pneumoniae -positive patients or the sera from healthy people. These findings will help to clarify the mimic epitopes of MgPa to facilitate diagnosis of the antigen and to understand the antigenic structure of MgPa.

BCRs of Mutated and Unmutated CLL Bind Peptides with Distinct Sequence Features: Evidence from Phage Display Libraries.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1118-1118
Author(s):  
Manuela Woelfle ◽  
Till Seiler ◽  
Rosa Catera ◽  
Hartmut Dohner ◽  
Stephan Stilgenbauer ◽  
...  
Keyword(s):  
Amino Acid ◽  
Phage Display ◽  
Synthetic Peptides ◽  
Specific Binding ◽  
Phage Display Library ◽  
Amino Acid Sequences ◽  
Phage Display Technology ◽  
Ligand Interactions ◽  
Distinct Sequence ◽  
Peptide Phage Display

Abstract In CLL, the use of specific IgV genes to code the clone’s BCR is non-random and there is an apparent selection for particular genetic and amino acid structures that can be shared by different patients, supporting the hypothesis that antigenic stimulation influences the development and course of CLL. As the binding specificities of the BCR are largely unknown, a vast variety of antigens may affect the BCRs and defined antigens have yet to be identified. Therefore, we used peptide phage display technology to identify ligands for CLL BCRs. BCRs from 2 IgVH unmutated (U-CLL) and 3 mutated (M-CLL) patients were expressed as IgG1 mAbs and used to probe a 12-mer peptide phage display library. In each case, after 3 rounds of selection, we isolated ligands reactive with the CLL mAbs. For the 3 M-CLL mAbs, phage clones carrying peptide inserts with conserved consensus motifs were found. Specificity of the BCR-ligand interactions was demonstrated in direct and indirect ELISA, since selected phage clones and synthetic peptides bound to their respective M-CLL mAb but not to other M-CLL mAbs. Variation of the amino acid sequence of the synthetic peptides significantly altered their reactivity with the corresponding M-CLL mAb. Furthermore, synthetic peptides were bound only by the proper mAb/BCR, but not by mAbs of other M-CLL or U-CLL patients with BCRs comprised of different IgVH genes, supporting the hypothesis that BCRs of M-CLL recognize a defined epitope. In contrast, the mAbs from 2 U-CLL cases did not select phages bearing a consensus motif. Rather these U-CLL mAbs bound multiple phages expressing the same 12-mer peptides, although these differed in sequence between the two U-CLL cases tested. Furthermore, 2 separate selection procedures using 1 U-CLL mAb isolated multiple phage bearing the same 12-mer sequence on each occasion as well as another set of phages with a completely distinct sequence in 1 of 2 selections. ELISAs demonstrated specific binding of all phage clones and of the synthetic peptides by the U-CLL mAbs. Despite this level of specificity, the 2 U-CLL mAbs also reacted with peptides isolated from panning with other CLL mAbs, thereby displaying considerable polyreactivity. Rather than binding only one distinct epitope, mAbs from U-CLL appear capable of interacting with multiple, unrelated structures. Finally, one of the peptides isolated with an U-CLL mAb was bound by all of the CLL mAbs tested, including those from M-CLL cases; therefore this target is antigenically “polyreactive”. Thus, phage display is a feasible approach to identify specific ligands for CLL BCRs. The two classes of BCRs in M-CLL and U-CLL show substantially different binding properties - the former binding shared amino acid motifs and the latter binding multiple ligands of distinct and identical 12-mer amino acid sequences. These peptides can be used to analyze more precisely the binding sites of CLL BCRs as well as the consequences that ensue after BCR crosslinking, and they might help develop BCR-specific therapeutic agents.

scholarly journals Factor VIII Epitope Analysis Using a Random Peptide Phage-Display Library Approach in the Sippet Cohort

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3176-3176
Author(s):  
Shermarke Hassan ◽  
Guido Baselli ◽  
Kaia Palm ◽  
Frits Richard Rosendaal ◽  
Roberta Palla ◽  
...  
Keyword(s):  
Phage Display ◽  
Factor Viii ◽  
Epitope Mapping ◽  
Hemophilia A ◽  
Phage Display Library ◽  
Mapping Technique ◽  
Discriminative Performance ◽  
Inhibitor Development ◽  
Random Peptide ◽  
Peptide Phage Display

Abstract Background Inhibitor development is the most severe complication of hemophilia A care, and is associated with increased morbidity and mortality. Aims The aim of this study was to use a novel epitope mapping method to explore the factor VIII (FVIII)-specific epitope profile in the SIPPET cohort population. Methods The population consisted of 122 previously untreated patients with severe hemophilia A that were followed-up for 50 days of exposure to FVIII. Sampling was performed before FVIII treatment and at the end of the follow-up. The outcome was inhibitor development. The FVIII-specific IgG epitope repertoire was assessed by means of a novel high-throughput epitope mapping technique using a random peptide phage-display library. Using this assay, a set of affinity-selected 12-mer peptide sequences (also called mimotopes) that were strongly bound by FVIII-specific antibodies were identified. These mimotopes were clustered on the basis of sequence similarity and a consensus motif was generated for each mimotope cluster. Discriminative performance of these mimotope clusters was assessed by ROC analysis. Mimotope clusters were mapped onto the 3D structure of a B-domain deleted FVIII model using a B-cell epitope prediction algorithm (Mapitope). Results The FVIII-specific antibody response is polyclonal with several mimotope clusters. The most predominant mimotope clusters in inhibitor patients were mapped to the heavy chain of the FVIII molecule. Using plasma samples taken before exposure to FVIII, three mimotopes (with the consensus motifs "QM", "PSLxWK" and "SWPHxxxxK") were identified that predicted inhibitor development (with an AUC of 0.76, 0.80 and 0.76 respectively). Conclusion Information on immunodominant epitope clusters can be used to generate novel, less immunogenic FVIII proteins and set up diagnostic tests that predict the risk of inhibitor development before starting treatment with FVIII. Figure 1 Figure 1. Disclosures Palm: Protobios LLC: Current Employment, Patents & Royalties: Inventor of the patent application (PCT Application No. US/14079626) filed by Protobios that covers the use of phage display method to manipulate and monitor humoral immunity.. Palla: Pfizer: Other: Travel support; Kedrion: Other: Travel support; Novonordisk: Speakers Bureau. Peyvandi: Roche: Consultancy, Honoraria; Sanofi: Consultancy, Honoraria; Sobi: Consultancy, Honoraria; Takeda: Consultancy, Honoraria.

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

2003 ◽  
Vol 307 (4) ◽  
pp. 791-796 ◽  
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

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

2017 ◽  
Vol 7 (1) ◽  
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

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

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

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

1997 ◽  
Vol 270 (1) ◽  
pp. 26-35 ◽  
Author(s):  
Shane Atwell ◽  
John B.B Ridgway ◽  
James A Wells ◽  
Paul Carter
Keyword(s):  
Phage Display ◽  
Phage Display Library

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

1994 ◽  
Vol 124 (3) ◽  
pp. 373-380 ◽  
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.

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

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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