Simplified Protein Purification with CLīM Technology

Achieving protein purification excellence has never been easier with TriAltus' CLīM technology.

 

 Here are the straightforward steps:

  1. Bind: Apply your lysate containing the CL-7 tagged protein to our Im7 Resin-packed column. Im7 Resin, featuring specific affinity for the CL7-tag, ensures minimal off-target binding. The high salt tolerance of the resin allows for reduced nonspecific interactions through the ability to load and wash in high salt buffers.

 

  1. Wash: Conduct 2-3 cycles of column washing with an alternation of low and high salt buffer. This step eliminates impurities and readies the column for elution.

 

  1. Elute: Utilize our highly efficient and site-specific elution proteases to cleave the CL7 tag:
    • SUMO Protease: This site-specific protease cleaves after the SUMO-domain, typically positioned N-terminal to the target protein. It allows for the generation of the native N-terminus.
    • PreScission Protease: This protease recognizes and cleaves at a specific cleavage site sequence. Usable at either the N or C terminal to the target protein, PreScisson protease leaves only a minimal number of residual amino acids.

Both proteases are engineered for exceptional purity, speed, and efficiency. Many clients find that the minimal amount of co-eluting protease is inconsequential since the purified protein already meets their application's purity requirements.

Alternatively, a tandem affinity column can be used to capture the co-eluted protease passively as it elutes from the Im7 column. 

 

  1. Purity Check: Confirm your eluted protein's purity using standard quantitation and purity analyses. CLīM's salt-independent, specific binding guarantees a clean, pure final product.

At TriAltus, we maintain rigorous standards for assessing protein purity through two distinct methods:

    • A260/A280 Ratio: This ratio provides valuable insights into the extent of nucleic acid contamination. You can delve deeper into this topic by referring to our earlier blog post linked here (https://bit.ly/3u80L2h). It's worth noting that CLīM consistently delivers proteins that attain optimal A260/A280 ratios.
    • SDS-PAGE Analysis: Our thorough analysis includes densitometry of a Coomassie-stained protein gel, providing an accurate evaluation of protein contaminants within our final protein product. With CLīM, we consistently achieve protein purities exceeding 95%.

These stringent assessments ensure that our protein purification processes with CLīM consistently yield high-quality and pure protein products.

 

TriAltus Products Needed: To implement the CLīM purification method with your CL7-tagged target protein, you'll require essential TriAltus products:

  • Im7 Resin for binding (https://bit.ly/49uuQYK)
  • highly active elution proteases (SUMO P or PSC P). (https://bit.ly/42ASFvL)

 

Purification protocols are also available here: (https://bit.ly/42G5d4R)

 

Why Choose CLīM: By opting for TriAltus' CLīM technology, you're not just purifying proteins; you're embracing a streamlined, cost-effective, high-yield method that surpasses traditional systems. Our robust one-step chromatography guarantees exceptional results.

 

Custom Purification Services: Unburden your team by entrusting us with your critical protein purification projects. Our custom protein purification service, powered by CLīM technology, ensures unparalleled purity and efficiency. Let us handle these tasks, allowing your team to focus on essential projects without compromising quality. We are equipped to deliver proteins in compliance with ISO 13485 standards, meeting the highest quality and regulatory requirements. Partner with TriAltus to accelerate your research while maintaining uncompromised excellence. (https://bit.ly/3HOYFrh)

 

Contact Us: For personalized consultations and to experience the innovation that will redefine your protein purification journey, reach out to our CLīM experts:

Email: info@trialtusbioscience.com Phone: +1.205.453.8242

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