An integrated microfluidic system for screening of peptides specific to colon cancer cells and colon cancer stem cells using the phage display technology

Jui, Yu; Wu, Huei Wen; Hung, Lein Yu; Chang, Hwan‐You; Wang, Kuan; Lee, Gwo‐Bin

doi:10.1109/nems.2014.6908845

The 9th IEEE International Conference on Nano/Micro Engineered and Molecular Systems (NEMS) 2014

DOI: 10.1109/nems.2014.6908845

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An integrated microfluidic system for screening of peptides specific to colon cancer cells and colon cancer stem cells using the phage display technology

Yu Jui¹,

Huei Wen Wu²,

Lein Yu Hung³

et al.

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Cited by 2 publications

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Continuous microfluidic assortment of interactive ligands (CMAIL)

Hsiao

Huang

et al. 2016

Sci Rep

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Finding an interactive ligand-receptor pair is crucial to many applications, including the development of monoclonal antibodies. Biopanning, a commonly used technique for affinity screening, involves a series of washing steps and is lengthy and tedious. Here we present an approach termed continuous microfluidic assortment of interactive ligands, or CMAIL, for the screening and sorting of antigen-binding single-chain variable antibody fragments (scFv) displayed on bacteriophages (phages). Phages carrying native negative charges on their coat proteins were electrophoresed through a hydrogel matrix functionalized with target antigens under two alternating orthogonal electric fields. During the weak horizontal electric field phase, phages were differentially swept laterally depending on their affinity for the antigen, and all phages were electrophoresed down to be collected during the strong vertical electric field phase. Phages of different affinity were spatially separated, allowing the continuous operation. More than 105 CFU (colony forming unit) antigen-interacting phages were isolated with ~100% specificity from a phage library containing 3 × 109 individual members within 40 minutes of sorting using CMAIL. CMAIL is rapid, sensitive, specific, and does not employ washing, elution or magnetic beads. In conclusion, we have developed an efficient and cost-effective method for isolating and sorting affinity reagents involving phage display.

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Continuous microfluidic assortment of interactive ligands (CMAIL)

Hsiao

Huang

et al. 2016

Sci Rep

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Design, Screening, and Testing of Non-Rational Peptide Libraries with Antimicrobial Activity: In Silico and Experimental Approaches

et al. 2020

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One of the challenges of modern biotechnology is to find new routes to mitigate the resistance to conventional antibiotics. Antimicrobial peptides (AMPs) are an alternative type of biomolecules, naturally present in a wide variety of organisms, with the capacity to overcome the current microorganism resistance threat. Here, we reviewed our recent efforts to develop a new library of non-rationally produced AMPs that relies on bacterial genome inherent diversity and compared it with rationally designed libraries. Our approach is based on a four-stage workflow process that incorporates the interplay of recent developments in four major emerging technologies: artificial intelligence, molecular dynamics, surface-display in microorganisms, and microfluidics. Implementing this framework is challenging because to obtain reliable results, the in silico algorithms to search for candidate AMPs need to overcome issues of the state-of-the-art approaches that limit the possibilities for multi-space data distribution analyses in extremely large databases. We expect to tackle this challenge by using a recently developed classification algorithm based on deep learning models that rely on convolutional layers and gated recurrent units. This will be complemented by carefully tailored molecular dynamics simulations to elucidate specific interactions with lipid bilayers. Candidate AMPs will be recombinantly-expressed on the surface of microorganisms for further screening via different droplet-based microfluidic-based strategies to identify AMPs with the desired lytic abilities. We believe that the proposed approach opens opportunities for searching and screening bioactive peptides for other applications.

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An integrated microfluidic system for screening of peptides specific to colon cancer cells and colon cancer stem cells using the phage display technology

Cited by 2 publications

References 8 publications

Continuous microfluidic assortment of interactive ligands (CMAIL)

Continuous microfluidic assortment of interactive ligands (CMAIL)

Design, Screening, and Testing of Non-Rational Peptide Libraries with Antimicrobial Activity: In Silico and Experimental Approaches

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