BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3628

Peptides:

Sequences	Affinity Values
ACVWWDHTYC(8/19) ACNWWDLTLC(6/19) ACTWWDMAFC(4/19) ACEWWDVTYC(1/19)	[2.0156±0.0932] [1.8097±0.0919] [1.8452±0.1287] [2.5513±0.0711]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 37126866

Target Name:

Anti-CPA monoclonal antibody MAB-1418

Template Name:

Cyclopiazonic acid, CPA

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:

ELISA

Affinity Measurement Description:

The analytical signal was obtained by measuring the absorbance at 450 nm with a CLARIOstar microplate reader from BMG (Ortenberg, Germany). Data shown were reproduced from Figure 2.

Brief Description:

A pre-selection step against BSA was carried out to eliminate non-specific phages prior to the actual selection with mAb-1418 in each round.

BiopanningDataSet Comments:

The mimotope sequence, ACNWWDLTLC, performed best in phage-based immunoassays, surface plasmon resonance binding analyses, and UC-RET-based immunoassays. To develop a homogeneous assay, upconversion nanoparticles (UCNP, type NaYF4:Yb3+, Er3+) were used as energy donors and coated with streptavidin to anchor the synthetic biotinylated mimotope. Alexa Fluor 555, used as an energy acceptor, was conjugated to the anti-CPA antibody fragment. The homogeneous single-step immunoassay could detect cyclopiazonic acid (CPA) in just 5 min and enabled a limit of detection (LOD) of 30 pg/mL (1.5 μg/kg) and an IC50 value of 0.36 ng/mL. No significant cross-reactivity was observed with other co-produced mycotoxins. Finally, we applied the novel method for the detection of CPA in spiked maize samples using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) as a reference method.

Curator:

Bowen Li

Curation Date:

2023-10-13

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