Laterally mobile, functionalized self-assembled monolayers at the fluorous-aqueous interface in a plug-based...

Area of Science:

• Interface science
• Microfluidics
• Nanotechnology

Background:

• Self-assembled monolayers (SAMs) are crucial for controlling interfaces in biological studies and nanotechnology.
• Current SAMs lack lateral mobility, and lipid-based methods are not ideal for high-throughput screening.
• A need exists for methods that combine the stability of SAMs with the mobility required for extensive experimental screening.

Purpose of the Study:

• To develop a novel method for generating functionalizable and mobile self-assembled monolayers (SAMs).
• To utilize microfluidic plugs for rapid exploration of chemical space in interface studies.
• To enhance protein crystallization screening through improved interface control.

Main Methods:

• Designed a new fluorinated amphiphile, RfNTA, incorporating his-tag binding chemistry.
• Synthesized RfNTA using an improved one-step method for RfOEG under Mitsunobu conditions.
• Employed plug-based microfluidics to generate mobile, functionalized SAMs at the fluorous-aqueous interface.

Main Results:

• RfNTA enabled specific protein binding and orientation at the interface, even with competing surfactants.
• Applied the method to crystallize a his-tagged membrane protein (Reaction Center from Rhodobacter sphaeroides).
• Achieved a significant increase in the range of crystal-producing conditions, success rate, nucleation rate, and crystal quality over 2400 trials.

Conclusions:

• The developed method provides a complementary approach to existing SAM and lipid techniques for interface manipulation.
• This plug-based microfluidic system facilitates rapid screening and optimization of conditions for protein crystallization.
• The functionalizable, mobile SAMs show promise for advancing studies in biological interfaces, heterogeneous reactions, and nanotechnology.