SOLUTIONS

ToyoScreen

ToyoScreen Columns 

In manufacturing, chromatography unit operations are some of the most critical in the purification of a new recombinant therapeutic protein. In many cases two to four chromatographic steps are needed to achieve the desired purity level for the target molecule. Each step can require up to multi-hundred and perhaps even thousands of liters of a specific resin for success.

The first part of the development process is the laboratory scale screening and selection of a resin. Due to the diversity in available ligand chemistries and base matrices offered by different vendors (e.g., agarose, methacrylate, styrene/divinylbenzene etc.) it is prudent at this stage to screen as many resins as possible. A thorough evaluation is a necessity as each target molecule has very different physical and chromatographic properties. Very often a resin that worked in the past for a similar molecule will not work as effectively for the new target molecule.


Historically, resin screening was accomplished by manually packing various bulk resins into small columns requiring a significant investment in time and cost. In order to improve the efficiency of resin screening experiments, pre-packed ToyoScreen columns containing the most popular Toyopearl resins, now provide a low cost and convenient alternative for the evaluation of Toyopearl ligand chemistries.


Screening of TOYOPEARL HIC Resins with Protein Standards 

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Resins:             A. TOYOPEARL Ether-650M B. TOYOPEARL PPG-600M
                          C. TOYOPEARL Phenyl-650M D. TOYOPEARL Butyl-650M
                          E. TOYOPEARL SuperButyl-550C F. TOYOPEARL Hexyl-650C
Column size:    ToyoScreen, 1 mL
Mobile phase:  Buffer A: 0.1 mol/L phosphate buffer + 1.8 mol/L sodium sulfate, pH 7.0 
                          Buffer B: 0.1 mol/L phosphate buffer, pH 7.0
Gradient:          30 min linear gradient buffer A to buffer B
Flow rate:         1 mL/min
Samples:          ribonuclease A, lysozyme, α-chymotrypsinogen,  1 mg/mL each 

 

Effect of eluent pH on chromatogram (separation of cell culture supernatant (Anti-TSH)) 

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Resin:               TOYOPEARL Phenyl-650M
Column size:   ToyoScreen, 1 mL
Mobile phase:  Buffer A: 0.1 mol/L phosphate buffer + 1.8 mol/L ammonium sulfate,
                          pH 7.0 Buffer B: 0.1 mol/L phosphate buffer, pH 7.0
Gradient:          30 min linear gradient, 30 CV
Flow rate:         1 mL/min
Injection vol.:  200 µL
Samples:          cell culture supernatant (× 4 diluted) (antibody: Anti-TSH) 

 

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