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Updated: Jul 7, 2026

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

Engineering peptide linkers for scFv immunosensors.

Zhihong Shen¹, Heping Yan, Ying Zhang

¹Department of Chemistry, Oakland University, Rochester, Michigan 48309, USA.

Analytical Chemistry

|February 23, 2008

Summary

Engineered single-chain fragment variable (scFv) antibodies with charged linkers show improved immobilization on self-assembled monolayers (SAMs). This strategy enhances detection sensitivity for biosensing applications.

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Area of Science:

Biomolecular Engineering
Surface Chemistry
Biosensor Development

Background:

Single-chain fragment variable (scFv) antibodies offer versatile platforms for biosensing.
Surface coupling chemistry provides adaptable methods for immobilizing biomolecules.
Self-assembled monolayers (SAMs) are functionalized surfaces used in various applications.

Purpose of the Study:

To engineer scFv linkers for enhanced immobilization on functionalized SAMs.
To investigate the impact of charged peptide linkers on scFv orientation and stability.
To improve the detection limits of scFv-based immunosensors.

Main Methods:

Engineering of a 15-mer peptide linker (RGRGRGRGRSRGGGS) incorporating six arginines into A10B scFv (A10B scFv-RG3).

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Last Updated: Jul 7, 2026

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

Utilizing anionic charged SAM templates, including 11-mercaptoundecanoic acid (MUA) and poly(sodium 4-styrenesulfonate) (PSS).

Comparing the performance of engineered scFv constructs with traditional antibody formats and other scFv variants.

Main Results:

Anionic SAM templates facilitated oriented immobilization and reduced protein denaturation of scFvs.
A 42-fold improvement in detection limits (to < 0.2 nM) was achieved with MUA/A10B scFv-RG3 compared to A10B Fab.
A picomolar detection limit was achieved using protein A-coated gold nanoparticles.

Conclusions:

Engineered scFv with charged linkers combined with SAM technology offers a robust strategy for biosensor development.
This approach significantly enhances sensitivity and specificity in scFv immunosensors.
Nanoparticle mass amplification further boosts the performance of these advanced immunosensing systems.