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Chromatography Media - IVD Core Raw Materials Supplier | Nanotein

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  • Time of issue:2022-07-07 04:42
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(Summary description)

Chromatography Media - IVD Core Raw Materials Supplier | Nanotein

(Summary description)

  • Categories:News
  • Author:
  • Origin:
  • Time of issue:2022-07-07 04:42
  • Views:
Information

Background

The pandemic has influenced the world extensively, especially in the biotech industry. Many IVD companies have found that the supply chain has become much more expensive and unstable compared to the pre-pandemic era. Therefore, finding alternative suppliers has become one of the most effective ways to increase supply chain stability.

 

About Nanotein

Nanotein is a subsidiary of the Genfine Group. Suzhou Nanotein Biotechnology Co., Ltd. is committed to the corporate culture of ‘customer-centric, technological innovation-oriented, and provides stable, reliable, cost-effective independent products and professional integrated solutions for the field of life sciences. Provide high-quality microsphere materials with precise and controllable size, material, structure and matrix for biomedicine, blood products, diagnosis, detection, and other fields, and strive to become the world's leading supplier of biological microsphere materials and services.

 

Key Raw Materials for Enzyme Production- Chromatography Media

 

Most recombinant raw enzymes will introduce protein tags to facilitate rapid downstream purification and production. His tag chromatography media has become the most commonly used affinity tag.

 

His-tagged chromatography medium

We provide Ni-NTA Starose 6 Fast Flow prepacked products in different specifications, including 1 mL prepacked columns, 5 mL prepacked columns, and 10-110 mL XPXK prepacked columns.

Features

 

The Ni-NTA Starose 6 Fast Flow medium is a highly cross-linked agarose matrix, which chelates Ni2+ after coupling with NTA ligands. Ni2+ has 6 chelation sites, of which 4 sites are used for binding to NTA ligands, and the remaining 2 sites are used for binding to His-tag proteins.

 

Features

● Fast flow

● high capacity

● High purity

● Very low nickel ion shedding during use

●Easy to scale up

 

Applications

His-tag protein purification was performed using StarBio Ni-NTA Starose 6 Fast Flow, and the following results were obtained:

 

● Good reproducibility of purification

● The effect of purifying tags of different lengths is better

● The effect of purifying proteins of different molecular weights is good

● The purification scale is easy to scale up (1 mL, 5 mL and 300 mL)

 

Case 1 (His)6-tag protein was purified 6 times repeatedly

 

Column: Ni-NTA Starose 6 Fast Flow Column

Sample: His-tag Pro-1 (Mr~25000) E. coli

Sample Size: 100ml

Buffer 1: 20m M PB, 500 mM NaCl, 20 mM Imidazole, pH7.4

Buffer 2: 20m M PB, 500 mM NaCl, 500 mM Imidazole, pH7.4

Speed: 1.0 ml/min

Figure 1. SDS-PAGE electrophoresis detection of 6 repeated purifications

Analysis: Using the same sample, 6 times of purification experiments were carried out under the same process, without the need for medium regeneration, cleaning or Ni2+ supplementation, with excellent purity and recovery reproducibility.

 

Case 2: (His)10-tag protein purification

 

Column: Ni-NTA Starose 6 Fast Flow 1ml Column

Sample: (His)10-tag Pro-2 (Mr~75000) E. coli

Sample Size: 5ml

Buffer 1: 20m M PB, 500 mM NaCl, 35 mM Imidazole, pH7.4

Buffer 2: 20m M PB, 500 mM NaCl, 500 mM Imidazole, pH7.4

Speed: 0.2 ml/min

Figure 2. Detection of (His)10-tag protein purification by SDS-PAGE electrophoresis (1: Protein Marker 2: Loading 3: Flow 4: Wash)

 

Case 3: Different molecular weight (His) 6-tag protein purification

Column: Ni-NTA Starose 6 Fast Flow 1ml Column

Sample: (His)-tag Pro-3 (Mr~41000) E. coli

       (His)-tag Pro-4 (Mr~68000) E. coli

       (His)-tag Pro-5 (Mr~75000) E. coli

       (His)-tag Pro-1 (Mr~25000) E. coli

Sample Size: 5ml

Buffer 1: 20m M PB, 500 mM NaCl, X mM Imidazole, pH7.4

Buffer 2: 20m M PB, 500 mM NaCl, 500 mM Imidazole, pH7.4

Speed: 0.2 ml/min

Figure 3. Different molecular weight (His)6-tag purification SDS-PAGE electrophoresis detection (1: Protein Marker 2: Loading 3: Flow 4: Wash)

 

Case 4: (His)6-tag protein purification and amplification

Column: Ni-NTA Starose 6 Fast Flow 1ml Column

       Ni-NTA Starose 6 Fast Flow 5ml Column

       Ni-NTA Starose 6 Fast Flow 50/20 300 ml

Sample: (His)-tag Pro-1 (Mr~25000) E. coli

Sample Size:  Ni-NTA Starose 6 Fast Flow 1ml Column 20mL

            Ni-NTA Starose 6 Fast Flow 5ml Column 100mL

            Ni-NTA Starose 6 Fast Flow 50/20 300 ml 8750mL

Buffer 1: 20m M PB, 500 mM NaCl, 20 mM Imidazole, pH7.4

Buffer 2: 20m M PB, 500 mM NaCl, 500 mM Imidazole, pH7.4

Speed: Ni-NTA Starose 6 Fast Flow 1ml Column 0.2mL/min

      Ni-NTA Starose 6 Fast Flow 5ml Column 1.0mL/min

      Ni-NTA Starose 6 Fast Flow 50/20 300 ml 40mL/min

Figure 4. (His)6-tag protein purification amplification detection by SDS-PAGE electrophoresis (1: Protein Marker 2: Loading 3: Flow 4: Wash)

Analysis: The purification scale of His-tag Pro-1 protein was enlarged from a 1 mL prepacked column to a 5 mL prepacked column, and then further enlarged to a 300 mL chromatography column. The purity of the target protein was 95.1%, 94.5% and 96.8%, respectively. The yields were 94.5%, 95.2% and 97.2%, respectively. Compared with the 1 mL prepacked column and the 5 mL prepacked column, the purification of the protein using the 300 mL column showed a slight increase in purity and yield, indicating that Ni-NTA Starose 6 Fast Flow can be used without affecting the purity and yield. The amplification of the purification process of His-tag-tagged proteins can be realized under the premise of high efficiency.

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