Background

Traditional protein purification, though widely used for its effectiveness in isolating proteins quickly from complex biological samples like cell extracts or culture media, can be detrimental to delicate proteins. The necessary elution step in affinity chromatography, which often involves altering the chemical environment through changes in pH, salt levels, or the introduction of competing molecules, can negatively impact the protein's structure (denaturation), cause it to clump together (aggregation), or render it non-functional (loss of activity). This highlights an increasing demand for purification techniques that are more efficient, gentler on the protein, and potentially more economical for obtaining pure and active proteins needed for research, diagnostic tools, and therapeutic applications.

Fig.1 Recombinant protein: a key component involved in various applications.^1,3

Light-based Antibody Purification Service at Creative Biolabs

To address the inherent limitations of traditional protein purification methods, Creative Biolabs offers cutting-edge and cost-effective light-based antibody purification service. Leveraging the innovative Azo-tag technology, our platform ensures more efficient and gentler protein purification.

We equip your target protein with an azobenzene tag, enabling specific binding to an α-cyclodextrin affinity column in its stable trans-configuration, facilitating thorough removal of unwanted contaminants through washing. Subsequently, a simple application of near-UV light triggers a reversible conformational change in the tag to the cis-configuration, dramatically reducing its affinity for the column matrix. This light-induced release allows for the elution of your purified protein in your buffer of choice, completely eliminating the need for harsh chemical eluents, salt gradients, or pH shifts. By harnessing the dynamic equilibrium between the light-responsive tags' isomers and their interaction with the affinity matrix, our platform provides a highly controlled and exceptionally gentle method for obtaining high-quality, functional proteins tailored to your specific research, diagnostic, or therapeutic needs. Partner with Creative Biolabs to revolutionize your protein purification workflow and unlock the full potential of your valuable protein targets.

Fig.2 Light-based protein purification principle.^2,3

Sample Types

Our service shows promise for a broad range of recombinant proteins that can be genetically modified to incorporate the Azo-tag, which include:

• Enzyme
• Antibody fragments
• Intact antibodies
• Sensitive proteins (prone to denaturation or loss of activity under traditional conditions)

Scientific Perspective

Highlights: This research introduces a novel light-controlled affinity chromatography technique for purifying recombinant proteins. The researchers demonstrated the effectiveness of single-step purification for purifying these tagged proteins in their functional state under native buffer conditions, even in a high-throughput manner.

Fig.3 Light-activated α-CD columns enable rapid, one-step purification of various Azo-tagged proteins from complex mixtures.^2,3

Fig.4 High-throughput, single-step purification of Azo-tagged proteins using a 96-well format.^2,3

Service Advantages

• Efficient single-step purification
• Gentle purification without chemical interactions
• Purified proteins with high activity
• Robust quality control system

Frequently Asked Question

Q1: What are some of the challenges involved?

A1: Navigating proteome complexity presents significant challenges, given the sheer scale of thousands of distinct proteins - many existing at trace concentrations or featuring post-translational modifications. Developing comprehensive antibody libraries demands meticulous engineering to achieve sufficient diversity for broad proteome coverage.

High-throughput workflows introduce data management hurdles, requiring robust systems to process and interpret massive screening outputs. Mitigating off-target interactions remains critical, as antibodies must demonstrate precise binding without cross-reactivity. Finally, scaling these efforts while balancing technical precision and cost-efficiency continues to test project viability. If you are experiencing issues such as poor protein purification efficiency, purified protein denaturation, or extended purification times, please feel free to contact us for further discussion.

References

1. Remans, Kim, et al. "Protein purification strategies must consider downstream applications and individual biological characteristics." Microbial Cell Factories 21.1 (2022): 52.
2. Mayrhofer, Peter, et al. "Protein purification with light via a genetically encoded azobenzene side chain." Nature Communications 15.1 (2024): 10693.
3. Distributed under Open Access License CC BY 4.0, without modification.

For research use only. Not intended for any clinical use.