Why optimize protein expression conditions in E. coli?

Optimization improves protein solubility, yield, and reduces impurities, ensuring reliable downstream use.

Why remove His-tags from recombinant proteins?

His-tag removal reduces potential interference in structural or functional studies and ensures native-like protein conformation.

How does periplasmic expression improve antibody fragment quality?

Directing antibody fragments to the periplasm promotes correct disulfide bond formation, yielding properly folded soluble proteins.

Can inclusion bodies be refolded into functional proteins?

Yes, refolding strategies can recover functional proteins from inclusion bodies, validated by QC assays.

What services does AtaGenix provide for protein expression?

AtaGenix provides end-to-end services from expression optimization, purification, tag removal, refolding, to QC validation across multiple platforms.

Protein Expression & Case Studies | AtaGenix Recombinant Protein Solutions

Release time: 2024-11-05 View volume: 520

Case 1:

1. Expression system optimization included host strains, induction temperature, induction time, inducer concentration, and culture media.

Host Strains	Induction Temperature	Induction Time	Inducer	Culture Media
T7E, BL21, C41, Arctic	16°C / 37°C	16h / 4h	IPTG	LB / Auto-induction medium

Fig. 1: Optimization of Protein Expression Conditions

Optimized conditions improved solubility and minimized impurities across different host strains.

2. 3C enzyme was used to remove the His-tag from the target protein. QC results are shown below:

Fig. 2: Removal of His-tag with 3C Enzyme Fig. 3: QC of His-tag Removed Protein

His-tag was efficiently cleaved, and QC confirmed target integrity with minimal by-products.

Case 2:

Antibody fragments expressed in E. coli often misfold due to disulfide bonds, forming inclusion bodies. By directing expression to the periplasmic space, properly folded soluble fragments were obtained.

1. Protein expression and QC results are shown below:

Periplasmic Expression and QC of Antibody Fragment

Fig. 4: Periplasmic Expression of Antibody Fragment Fig. 5: QC (Reduced / Non-reduced)

QC confirmed correctly folded soluble antibody fragments distinct from aggregated inclusion bodies.

Lane MW: Protein marker | Lane PP: Periplasmic Space

Case 3:

Many proteins in E. coli form inclusion bodies. To restore functionality, these were refolded into soluble proteins for activity assays.

1. Expression and QC results are shown below:

Optimization and Refolding QC of Target Protein

Fig. 6: Optimization of Target Protein Expression Fig. 7: Refolding QC of Target Protein

Refolding successfully converted insoluble inclusion bodies into soluble, functional proteins validated by QC bands.

Need optimized protein expression and validation support? AtaGenix provides end-to-end custom solutions for antibody fragments and recombinant proteins.

Contact AtaGenix

Support Center

Messages

By Method

By Application

Protein Expression & Case Studies | AtaGenix Recombinant Protein Solutions

About Us

Core Technologies

Resource Center