BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3226

Peptides:

Sequences	Affinity Values
FIPFDPMSMRWE SYIEPPERHRHR DDAKSRQGPLFR NVNEGKAGVTGW	[0.0057,NT] [0.0230,57] [0.0062,NT] [0.0036,NT]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 27989733

Target Name:

Myeloperoxidase

Template Name:

Anti-human myeloperoxidase antibody

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

Surface plasmon resonance (SPR),Quartz crystal microbalance (QCM)

Affinity Measurement Description:

A quartz crystal microbalance sensor (QCM D-300; Q-sense, Sweden) equipped with a QSX 301 gold chip was used to determine the molar binding ratio of hMPO to the immobilized ligand, i.e. the anti-hMPO antibody or the synthesized peptide. To study the binding kinetics and equilibrium, a surface plasmon resonance (SPR) biosensor (Biacore 3000, GE Healthcare, USA) equipped with a CM5 Sensor chip was used. The first colum of the affinity values represents the molar binding ratio determined by QCM, the second column denotes the Kd values (nM) determined by SPR.

Brief Description:

Before proceeding to the next round of biopanning, non-specific phages that were included in the selected phages were removed by a ‘subtraction’ (or negative selection) step by using epoxy-functionalized magnetic particles without a myeloperoxidase from human leukocytes (hMPO) coating (‘nude’ magnetic particles). The supernatant from this subtraction step was introduced to the next round.

BiopanningDataSet Comments:

We selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0.023, approximately 43-fold lower than that of the anti-hMPO antibody. The dissociation constant (Kd) was 57 nM, which was comparable to that of the native antibody (83 nM). We biotinylated the peptide at its N-terminus and attached the biotinylated peptide to the surface of streptavidin-coated magnetic particles to assess its ability to selectively capture hMPO. The binding equilibrium data were similar to the previous analyses; specifically, around 0.021 mol peptide bound to 1 mol of hMPO. Antigen capture was found to be selective and to be relatively little influenced by the presence of human serum albumin (HSA), an abundant constituent of serum.

Curator:

Bifang He

Curation Date:

2017-10-24

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