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A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells

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Microtitre Plate Based Cell-SELEX Method.

Munish Shorie¹, Harmanjit Kaur¹

¹Institute of Nano Science and Technology, Mohali-160062, India.

|September 17, 2021

Summary

We developed an efficient Cell-SELEX method using microtiter plates for isolating aptamers against E. coli. This optimized protocol accelerates the generation of high-affinity DNA aptamers for biosensing and therapeutic applications.

Keywords:

Aptamers Bacteria E. coli Gram negative bacteria Microbial detection SELEX bio-receptor screening

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Area of Science:

Biotechnology
Molecular Biology
Bioreceptor Development

Background:

Aptamers are versatile bioreceptors with applications in biosensing and therapeutics.
Systematic Evolution of Ligands by Exponential Enrichment (SELEX) is a key method for aptamer discovery.
Existing SELEX methods can be time-consuming and require optimization.

Purpose of the Study:

To develop a facile and efficient microtiter plate-based Cell-SELEX method.
To generate high-affinity aptamers against the gram-negative bacterium E. coli.
To reduce the number of SELEX rounds required for aptamer generation.

Main Methods:

A microtiter plate-based Cell-SELEX protocol was designed and optimized.
The method utilizes increased surface area and longer retention times for enhanced binding.
The protocol was applied for the selection of aptamers against E. coli.

Main Results:

The optimized Cell-SELEX protocol significantly reduced the number of SELEX rounds needed.
The method demonstrated efficiency in generating aptamers against E. coli.
The protocol offers potential for modification to target other cells and molecules.

Conclusions:

A facile microtiter plate-based Cell-SELEX method provides an efficient approach for aptamer generation.
This method accelerates the development of high-affinity bioreceptors for various applications.
The protocol is adaptable for selecting aptamers against diverse targets including prokaryotic and eukaryotic cells and glycan moieties.