BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3438

Peptides:

Sequences
CYLLCISPC CGSRCYPRC

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

ABI 3130

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 30198459

Target Name:

IgG from tegumentary leishmaniasis (TL) patients

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-C7C phage display library (CX7C)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Briefly, viral particles of the bacteriophage library (Ph.D.®-C7C library, New England BioLabs, Ipswich, MA, USA) were diluted in 250 μL of 50mM Tris-HCl pH 7.5, 150 mM NaCl and 0.1% Tween 20 buffer (TBS-T). The mixture was incubated for 30 min, at room temperature, with microspheres coupled to the IgG antibodies purified from healthy subjects for subtraction, which were magnetically captured. The remaining phages in the supernatant were recovered and transferred to a new tube, which was subjected to positive selection using IgG from TL patients. The supernatant was removed and the bound phages were washed five times in 1 mL TBS-T buffer, and were eluted in 500 μL of 0.2 M glycine buffer, pH 2.0. Next, 75 μL of 1 M Tris-base pH 9.0 were added to neutralize the acidic pH of the solution.

BiopanningDataSet Comments:

Two clones, namely A4 (CYLLCISPC) and A8 (CGSRCYPRC), were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls.

Curator:

Bifang He

Curation Date:

2019-05-28

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