Nuvia™ cPrime™ Media

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Nuvia™ cPrime™ Media

Overview

General Approach to Method Development Using Nuvia™ cPrime™ Media

Developing an effective and robust method with Nuvia cPrime media is simple. Below is general information about the binding and elution mechanisms and an approach to guide method development; results will vary depending on the protein of interest and the feed composition.

The binding and elution mechanisms of Nuvia cPrime media are determined chiefly by pH and salt. The high salt tolerance of the media often allows for direct loading at high conductivity. An increase in pH will in most cases achieve elution. Conductivity is another way to achieve and/or optimize elution and the final method is often a combination of an increase in pH and/or an increase/decrease in salt concentration. In some cases, the use of an elution buffer modifier or a different salt in the elution buffer may be required for optimal elution, recovery, and resolution.

The schematic below outlines a recommended approach to method development. In most cases, conducting a few simple experimental designs to identify optimal binding and elution conditions will yield an effective, robust, and scalable method.

Simple Method Development

1. Load feed or eluate from previous step directly without dilution onto the Nuvia cPrime media column. To elute, use an increasing pH gradient. If satisfactory elution and recovery are achieved, refine and/or make a step gradient to complete the step (pH 4–8, depending on protein).

2. If elution is not satisfactory after step 1, run a salt gradient to disrupt electrostatic or hydrophobic interactions that may be preventing elution or broadening the peak. Use the pH where there was best elution (from step 1). The direction of this salt gradient (increasing or decreasing) can be easily assessed and will depend on the relative contributions of ionic vs. hydrophobic interactions involved in binding.

3. If elution is still unsatisfactory after step 2 of this process, consider using a modifier such as propylene glycol, urea, or arginine to disrupt any remaining interactions. Other modifiers may also be used; in some cases changing to another salt may also be required.

Learn more

Using High-Throughput (HT) Methods to Accelerate DOE

High-throughput experimentation (HTE) was used in this case study in concert with design of experiments (DOE) methodology to study the mass recovery of a basic therapeutic protein (pI 9.2–9.5) using a hydrophobic cation exchange mixed-mode resin.

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Nuvia cPrime hydrophobic cation exchange media are a new addition to Bio-Rad's family of mixed-mode purification products. These media are designed for process-scale purification of a wide variety of therapeutic proteins. Nuvia cPrime media's unique selectivity allows method developers to use hydrophobic and cation exchange interaction modes to achieve effective purification. More importantly, the media have a large design space for binding and elution, allowing for the development of highly robust methods in a commercial manufacturing setting. Nuvia cPrime media are built on a rigid, mechanically and chemically stable macroporous base matrix with a particle size optimized to deliver exceptional flow properties, fast mass transfer, and stability.

Nuvia cPrime delivers value by providing:

  • New and unique selectivity
  • Salt tolerant media
  • Simple method development
  • Large design space for capture and elution of a variety of biotherapeutic proteins
  • High recovery of target protein
  • Mechanical and chemical stability
Functional group Hydrophobic weak cation exchange
Base matrix composition Macroporous highly crosslinked polymer
Particle size 70 µm ± 10 µm
Dynamic binding capacity hlgG: >40 mg/ml (at 10% breakthrough, 300 cm/hr)
Lactoferrin: >60 mg/ml
Ligand density 55–75 µeq/ml
Recommended maximum linear flow rate 50–600 cm/hr
Pressure vs. flow performance Under 2 bar at a flow rate of 600 cm/hr
(20 cm × 20 cm packed bed; 1.17 comp. factor)
pH stability short-term 2–14
pH stability long-term 3–13
Chemical stability 1.0 N NaOH, 1.0 N HCl, 25% HOAc, 8 M urea, 6 M GuHCl, 6 M KSCN, 3 M NaCl, 1% Triton X-100, 2% SDS + 0.25 M NaCl, 20% EtOH, 70% EtOH, 30% IPA
Shipping solution 20% ethanol with 30 mM sodium sulfite
Storage conditions 20% ethanol
Shelf life in 20% ethanol 5 years at room temperature
Number Description Options
10023853 Instruction Manual, Nuvia™ cPrime™ Hydrophobic Cation Exchange Resin, Rev B Click to download
6242 Nuvia™ cPrime™ Hydrophobic Cation Exchange Resin Product Information Sheet, Rev C Click to download [ Add to Cart (Free) ]
6992 Single-Step Purification of Raw Starch Digesting α-Amylase Using Nuvia™ cPrime™ Hydrophobic Cation Exchange Resin Click to download
6418 Purification of Recombinant Proteins on Nuvia cPrime Hydrophobic Cation Exchange Resin: A Simple Approach to Method Development, Rev A Click to download [ Add to Cart (Free) ]
6982 Mixed-Mode Resins/Media Selection Card Click to download [ Add to Cart (Free) ]
6983 Native and Recombinant Protein Purification Resins/Media Selection Card Click to download
6786 Monoclonal Antibody Purification Resin Selection Card, Ver A Click to download
6789 IgG Polyclonal Antibody Purification Resin Selection Card, Ver A Click to download
6791 Virus Purification Resin Selection Card, Ver A Click to download
6792 Aggregate Purification Resin Selection Card, Ver A Click to download
6241 A Purification Strategy for Clinical-Grade Monoclonal Antibody Using Hydrophobic Cation Exchange Chromatography, Rev A Click to download [ Add to Cart (Free) ]
6719 Development of an Efficient Manufacturing Process for Adenovirus, Ver A Click to download
6720 Development of a cGMP-Ready Purification Process for Adenovirus Purification (Poster), Rev A Click to download
6875 Practical Guide: Selecting the Optimal Resins for Monoclonal Antibody Process Purification, Ver A Click to download
6810 Practical Guide: Selecting the Optimal Resins for the Process Purification of Native and Recombinant Proteins, Ver A Click to download
6813 Practical Guide: Selecting the Optimal Resins for Endotoxin Depletion in Process Purification, Ver A Click to download
6807 Practical Guide: Selecting the Optimal Resins for Adenovirus Process Purification, Ver A Click to download
6808 Practical Guide: Selecting the Optimal Resins for Aggregate Removal, Ver A Click to download
6790 Practical Guide: Selecting the Optimal Resins for Mammalian Virus Process Purification and Clearance, Ver A Click to download
6881 Practical Guide: Selecting the Optimal Resins for Removal of DNA Contamination during Process Purification, Ver A Click to download
6946 Monoclonal Antibody Purification: Polish Purification Resins Brochure, Ver A Click to download
6887 Quick Monoclonal Antibody Purification Strategy: Non-Affinity Based Capture Flier, Ver A Click to download
6880 Quick Virus Purification Strategy: Adenovirus Purification Flier, Ver A Click to download
6869 Nuvia™ Chromatography Resins Publications List, Ver A Click to download

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