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Tag-free Protein Preparation Services

Tag-free Protein Preparation Services

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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

ModalityRecombinant protein
Starting materialsGene sequences, recombinant plasmid, or engineered strains
DeliverablesPurify: ＞80%, ＞85%, ＞90% Quantity: 0.2~10 g (to be determined by the project)
Expression system and formationE. coli periplasmic secretion, soluble and inclusion body expression; Yeast intracellular or secretion expression
Analysis methodsUltraviolet (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 impuritiesEndotoxin (optional)

Services Details

ServicesService DetailsMinimal Timeline(working day)
Gene synthesisCodon optimization for specific expression host7~10 days
Gene synthesis
Plasmid/vector constructionCloning into plasmid/vector2 days
Plasmid transformation
Polymerase chain reaction (PCR) verification, restriction enzyme digestion, gene sequencing3 days
Protein expression in E. coliPlasmid transformation,2 days
Cultivation in shake flask3 days
7L-sacle fermentation, optimization4 days /batch
Tag-free protein purification, optimization5~10 days /batch
Testing by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
Protein expression in yeastYeast competent cells preparation4 days
Plasmid linearization and electro-transformation
7L-sacle fermentation, optimization5 days
Tag-free protein purification, optimization7 days /batch
Tag-free protein purification5~10 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.

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