Potential application of antibody-mimicking peptides identified by phage display in immuno-magnetic separation of an antigen

Hien, Thai Bao Dieu; Maeng, Joon-Ho; Lee, Byung Heon; Seong, Gi Hun; Choo, Jaebum; Lee, E.K.

doi:10.1016/j.jbiotec.2012.06.039

Cited by 6 publications

(1 citation statement)

References 23 publications

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“…A Ph.D.-12 TM phage display peptide library containing 1.5 × 10 13 pfu/mL and 2.7 × 10 9 transformants was used to screen for targeting ricin or RCA120. Slight modification was made according to the previous literature [ 35 , 36 ]. Briefly, ricin (10 µg/mL in 0.1 M NaHCO 3 , pH 8.6) and RCA120 (10 µg/mL in 0.1 M NaHCO 3 , pH 8.6) were added to 96-cell plates (100 μL/well), respectively.…”

Section: Methodsmentioning

confidence: 99%

In Silico–Ex Vitro Iteration Strategy for Affinity Maturation of Anti-Ricin Peptides and the SPR Biosensing Application

Yang

Wang

Liu

et al. 2023

Toxins

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The highly toxic plant toxin ricin is one of the most known threatening toxins. Accurate and sensitive biosensing methods for the first emergency response and intoxication treatment, are always pursued in the biodefense field. Screening affinity molecules is the fundamental mainstream approach for developing biosensing methods. Compared with common affinity molecules such as antibodies and oligonucleotide aptamers, peptides have great potential as biosensing modules with more accessible chemical synthesis capability and better batch-to-batch stability than antibodies, more abundant interaction sites, and robust sensing performance towards complex environments. However, anti-ricin peptides are so scant to be screened and discovered, and an advanced screening strategy is the utmost to tackle this issue. Here, we present a new in silico-in vitro iteration-assisted affinity maturation strategy of anti-ricin peptides. We first obtained affinity peptides targeting ricin through phage display with five panning rounds of “coating-elution-amplification-enrichment” procedures. The binding affinity and kinetic parameters characterized by surface plasmon resonance (SPR) showed that we had obtained four peptides owning dissociation constants (KD) around 2~35 μM, in which peptide PD-2-R5 has the lower KD of 4.7 μM and higher stable posture to interact with ricin. We then constructed a new strategy for affinity maturity, composing two rounds of in silico-in vitro iterations. Firstly, towards the single-site alanine scanning mutation peptide library, the molecular docking predictions match the SPR evaluation results well, laying a solid foundation for designing a full saturation mutated peptide library. Secondly, plenty of in silico saturation mutation prediction results guided the discovery of peptides PD2-R5-T3 and PD-2-R5-T4 with higher affinity from only a limited number of SPR evaluation experiments. Both evolved peptides had increased affinity by about 5~20 times, i.e., KD of 230 nM and 900 nM. A primary cellular toxicity assay indicated that both peptides could protect cells against ricin damage. We further established an SPR assay based on PD-2-R5-T3 and PD-2-R5-T4 elongated with an antifouling peptide linkage and achieved good linearity with a sensitivity of 1 nM and 0.5 nM, respectively. We hope this new affinity-mature strategy will find its favorable position in relevant peptide evolution, biosensing, and medical countermeasures for biotoxins to protect society’s security and human life better.

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Section: Methodsmentioning

confidence: 99%

In Silico–Ex Vitro Iteration Strategy for Affinity Maturation of Anti-Ricin Peptides and the SPR Biosensing Application

Yang

Wang

Liu

et al. 2023

Toxins

View full text Add to dashboard Cite

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A High-Throughput Magnetic Nanoparticle-Based Semi-Automated Antibody Phage Display Biopanning

Ch’ng

Ahmad

Konthur

et al. 2018

Methods in Molecular Biology

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Design of a core–shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples

Özalp

Bayramoğlu

Arıca

et al. 2013

Appl Microbiol Biotechnol

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We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core-shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) of target bacterial cells from others, antibodies for Escherichia coli and Salmonella enterica serovar Typhimurium cells were immobilized on the magnetic beads via glutaraldehyde coupling reaction. Our IMS system successfully separated Salmonella cells when the concentrations of target (i.e., Salmonella) and interfering (i.e., E. coli) cells were at the same level. Polymerase chain reaction (PCR) assays amplifying the rfb/rfbE region of the E. coli genome and a 647-bp fragment of the invA region of Salmonella were performed as the specific selection to accurately confirm the presence of E. coli and Salmonella, respectively. IMS and multiplex PCR methods can be used for specific and quantitative detection of pathogens from food samples. Thus, this study developed a reliable and direct system for rapid detection of Salmonella and E. coli in food samples. In addition, IMS method could be easily adapted to detect other pathogens by selecting the pertinent antibody.

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Potential application of antibody-mimicking peptides identified by phage display in immuno-magnetic separation of an antigen

Cited by 6 publications

References 23 publications

In Silico–Ex Vitro Iteration Strategy for Affinity Maturation of Anti-Ricin Peptides and the SPR Biosensing Application

In Silico–Ex Vitro Iteration Strategy for Affinity Maturation of Anti-Ricin Peptides and the SPR Biosensing Application

A High-Throughput Magnetic Nanoparticle-Based Semi-Automated Antibody Phage Display Biopanning

Design of a core–shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples

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