High-throughput screening platform for discovering acid and heat stable sweet proteins
A high-throughput cell-based screening platform for identifying novel sweet proteins as natural sugar alternatives. Uses FRET-based assays in HEK293 cells expressing hT1R2/T1R3 sweet taste receptors to screen plant protein extracts, followed by immunoprecipitation and mass spectrometry for ligand identification.
Overview
This platform offers a high-throughput screening approach for discovering novel sweet proteins that can serve as natural sugar alternatives. Sweet proteins provide sweetness potency thousands of times greater than sucrose without glycemic effects, making them attractive candidates for sugar substitution in food and beverage applications. The technology addresses the need for acid- and heat-stable sweet proteins suitable for diverse formulation requirements.
The approach leverages the human sweet taste receptor complex (hT1R2/T1R3) found in taste buds to identify proteins that activate sweetness perception. Because sweet proteins do not share structural similarities yet all interact with this same receptor complex, a receptor-based screening strategy enables detection of structurally diverse candidates from crude plant extracts.
Technical specifications
Screening methodology:
- HEK293 cells are engineered to coexpress fusion proteins of hT1R2 and hT1R3 conjugated with GFP and RFP fluorescent tags
- FRET (Fluorescence Resonance Energy Transfer) signals detect ligand-induced dimer stabilization of the receptor complex
- FRET changes occur only when the receptor subunits come into close proximity through ligand binding, enabling reliable detection of sweet protein activity
- The cell model enables rapid screening of crude plant protein extracts in the absence of glucose
Ligand identification workflow:
- HEK293 cells expressing his-tagged hT1R2-P2A-hT1R3 are exposed to protein matrices containing candidate ligands
- Brazzein and Curculin serve as positive controls for assay validation
- Ligand-receptor complexes are cross-linked using reversible N-hydroxysuccinimide (NHS) esters
- Immunoprecipitation with Ni-NTA magnetic beads isolates complexes
- SDS-PAGE separation followed by MALDI-TOF mass spectrometry identifies unknown interacting partners
Technology readiness level
The platform is currently at an early research and development stage. The high-throughput FRET-based screening assay and ligand identification workflow using immunoprecipitation coupled with mass spectrometry have been designed but not yet experimentally validated. Future validation will involve testing the cell-based FRET assay with known sweet proteins and screening crude plant extracts to identify novel candidates with acid and heat stability properties suitable for food and beverage applications.
About pharmaplanter technologies inc
PharmaPlanter Technologies is a platform biotechnology company based in Burnaby, British Columbia, focused on translating early-stage scientific discoveries into viable biopharmaceutical products. The company develops integrated platforms including recombinant protein development, iPSC organoid culture, gene editing, and a proprietary gas vesicle (GV) nanoparticle scaffold for therapeutic delivery. By bridging discovery and development through specialized R&D, CRO services, and collaborative growth initiatives, PharmaPlanter aims to support emerging biotech ventures and advance translational innovation. Their technology suite is designed to integrate advanced therapeutic modeling with novel delivery methods, such as mucosal vaccination and personalized cancer immunotherapy.
The company addresses significant medical needs across areas like microbiome restoration, mucosal immunity, oncology, and metabolic regulation. Their clinical and preclinical pipeline includes oral fecal microbiota transplantation for gastrointestinal disorders, nasal gas vesicle-based vaccines for respiratory viruses, and intradermal cancer immunotherapies. By providing the infrastructure and scientific expertise necessary for complex drug development, PharmaPlanter enables its partners and clients to navigate the transition from scientific theory to clinical application, utilizing targeted delivery kinetics to optimize immune responses and therapeutic efficacy.
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