High-throughput clone screening followed by protein expression cross-check: A visual assay platform

Area of Science:

• Biotechnology
• Structural Biology
• Nanotechnology

Background:

• High-throughput molecular cloning is crucial for recombinant protein production in biotechnology and structural biology.
• Current methods for clone screening and expression verification are often time-consuming, costly, or complex.
• A need exists for rapid, cost-effective, and user-friendly protocols for both clone screening and protein expression confirmation.

Purpose of the Study:

• To develop and present a novel, visual method for high-throughput clone screening.
• To establish a cross-check mechanism for confirming the expression of desired recombinant proteins.
• To offer a cost-effective and sensitive alternative to traditional gel- and PCR-based techniques.

Main Methods:

• Utilized gold nanoparticle-based plasmonic detection for visual analysis.
• Developed a non-gel, non-PCR based protocol for clone screening.
• Integrated clone screening with protein expression verification in a single workflow.

Main Results:

• Demonstrated a visual detection technique for clone screening.
• Successfully cross-checked the expression of desired proteins.
• Achieved simultaneous high-throughput clone screening and expression determination.
• The protocol is characterized as easy, fast, sensitive, and inexpensive.

Conclusions:

• The gold nanoparticle-based plasmonic detection offers an efficient and accessible method for molecular cloning workflows.
• This technique significantly simplifies and accelerates the process of identifying successful clones and confirming protein expression.
• It provides a valuable tool for researchers in biotechnology and structural biology seeking to optimize recombinant protein production.