All Categories
ENEN
Tag-free Protein Preparation Services

Tag-free Protein Preparation Services

Home >  Tag-free Protein Preparation Services

Application

Tag-free Protein Preparation Services

Protein tags, as fusion partners used in recombinant DNA technologies, enable effective, single-step affinity purification for various proteins/peptides with different characteristics. Despite this important advantage, tags as exogenous amino acids may alter the protein structure and biological functions. Therefore, non-tagged (tag-free) proteins or peptides are more preferred to ensure safety and efficacy, especially in the biopharmaceutical field. A recombinant protein with a fusion tag has limited possibilities for obtaining authorization from medical/drug administration agencies.

Based on the microbial biologics R&D platform, Yaohai Bio-Pharma offers customized non-tagged (Tag-free) protein preparation services dedicated to meeting unique objectives for research, development, and manufacturing for clinical use.

Keywords: tag-free protein production, non-tagged protein purification, free tag-based protein preparation, tagless protein expression and purification, highly tag-free target protein Application: human medicine research, animal medicine development, recombinant vaccine development, recombinant large molecule biologics production


Our Microbial R&D Platform

We are experienced in microbial strain engineering, fermentation, and product purification.

Bacteria: Escherichia coli (E. coli)

Yeast: Pichia pastoris (P. pastoris), Saccharomyces cerevisiae (S. cerevisiae), Hansenula polymorpha (H. polymorpha)

Customized Options

Modality Recombinant protein
Starting materials Gene sequences, recombinant plasmid, or engineered strains
Deliverables Purify: ＞80%, ＞85%, ＞90% Quantity: 0.2~10 g (to be determined by the project)
Expression system and formation E. coli periplasmic secretion, soluble and inclusion body expression; Yeast intracellular or secretion expression
Analysis methods Ultraviolet (UV), bicinchoninic acid assay (BCA), Bradford, or Lowry for QuantitySodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for purify testingOptional: Western blot (WB）, enzyme-linked immunosorbent assay (ELISA), high performance liquid chromatography (HPLC)
Residual impurities Endotoxin (optional)

Services Details

Services Service Details Minimal Timeline(working day)
Gene synthesis Codon optimization for specific expression host 7~10 days
Gene synthesis
Plasmid/vector construction Cloning into plasmid/vector 2 days
Plasmid transformation
Polymerase chain reaction (PCR) verification, restriction enzyme digestion, gene sequencing 3 days
Protein expression in E. coli Plasmid transformation, 2 days
Cultivation in shake flask 3 days
7L-sacle fermentation, optimization 4 days /batch
Tag-free protein purification, optimization 5~10 days /batch
Testing by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
Protein expression in yeast Yeast competent cells preparation 4 days
Plasmid linearization and electro-transformation
7L-sacle fermentation, optimization 5 days
Tag-free protein purification, optimization 7 days /batch
Tag-free protein purification 5~10 days /batch
QC by SDS-PAGE

Additional Services (optional)

Fermentation Process Development

Tag-free Protein Purification Process Development

Pilot Scale up and GMP Manufacturing

Analysis and Testing

Our Features

We have worked on more than 300 engineered microbial strains, including E. coli DH5α, TOP10, Trans10, BL21; P. pastoris SMD1168H, X-33, GS115; PichiaPink strain1/2/3/4; and H. polymorpha.

We are experienced with E. coli periplasmic secretion, soluble and inclusion body expression, as well as yeast intracellular or secretion expression.

We establish a high-throughput screening platform to screen high-producing and stable strains from over one hundred candidate strains.

Get a Free Quote

Get in touch