Cellufine Resin Application List
At this location on the JNC web site are a range of Application Technical Notes describing recent purification of and polishing of monoclonal antibodies and whole viral vaccine particles. These technical articles are useful as a resource for development of your downstream process workflow with Cellufine products.
Purification of mAb from CHO culture supernatant with Cellufine products: Three step purification workflow
mAb’s derived from CHO cell culture supernatant can be purified with a conventional three step chromatography workflow. Initial mAb capture with Cellufine SPA-HC is applied followed by polishing with Cellufine MAX Q-h and Cellufine MAX GS.
Efficient mAb purification process using Cellufine chromatography media: Direct coupling of two polishing workflow steps (FT-FT mode)
In most downstream monoclonal antibody (mAb) purification processes, initial Protein A capture is usually followed by two polishing steps such as AEX followed by CEX. In this report, we have demonstrated an improved polishing workflow by directly connecting two flow-through chromatography steps (FT-FT mode): Cellufine MAX IB mixed mode and Cellufine GS cation exchange.
Two step purification of mAb with Cellufine rProA Affinity and Mixed Mode Chromatography Resins
In Technical Note, we introduce a two steps purification method using rProtein A chromatography for mAb capture and Cellufine MAX IB mixed mode as a second step polishing process. By efficiently combining capture and polishing it is possible to purify a mAb in a cost-effective workflow with minimal time spent moving between the two chromatography modes.
Purification of Human Coronavirus hOC43 with Cellufine Sulfate
Cellufine Sulfate has been widely used in virus purification. βcoronavirus hCoV OC43 has been purified with Cellufine Sulfate and is described in this Technical Note. This pseudo affinity chromatography resin has proved to be highly effective in the purification of βcoronavirus.
Two-step purification of T7 RNA Polymerase with Cellufine MAX DEAE and Phosphate Affinity Chromatography Resins
T7 RNA polymerase is an enzyme derived from T7 phage used to produce mRNA transcripts from the template DNA. In the manufacture of COVID-19 mRNA vaccines. In this report, we will introduce an example of purification of T7 RNA polymerase. T7 RNA polymerase was expressed in Escherichia coli (pAR1219) and then purifying the fermented solution with Cellufine Phosphate and Cellufine MAX DEAE (weak anion exchanger) to high purity. High-purity T7 RNA polymerase was able to be purified with Cellufine Phosphate, which is an affinity chromatography resin.
Purification of Capsular Polysaccharides of Streptococcus pneumoniae Serotype 19F by Cellufine
Streptococcus pneumoniae is one of the major pathogens causing high level morbidity and mortality worldwide, especially children and the elderly populations. Pneumococcal vaccines based on the capsular polysaccharide (CPS) on the bacteria surface, which is one of the most important virulence factors, have been used to prevent these infectious diseases. Traditional ethanol precipitations are general processes for purifying CPSs, which are suitable for most serotypes. However, they are complex and time consuming. As a result, pneumococcal vaccines are quite expensive. Here we would like to propose simplified two step chromatography purification process of CPS without ethanol precipitations. This process includes hydrophobic interaction chromatography (HIC) and anion exchange chromatography (AEX). We introduce two optimized resins, Cellufine MAX Butyl HS (HIC) and Cellufine MAX Q-hv (AEX), for this process.
An easy-to-understand explanation of antibody drug manufacturing methods
Antibody drugs have been attracting attention in the medical field in recent years because of their high specificity and effectiveness against specific pathogens and diseases. However, their manufacturing method is very complex and requires advanced technology. This time, we will explain the manufacturing process of antibody drugs in an easy-to-understand manner..
Separation of complete and empty AAV particles using AEX chromatography media based on cellulose-based MLP
To address the challenge of removing empty particles (i.e., DNA-unencapsulated particles) in AAV production, we developed and evaluated an AEX chromatography resin based on cellulose-derived monolithic particles (MLP) with continuous pores. By controlling particle size and pore size, we achieved improved dynamic binding capacity and resolution compared to conventional AEX chromatography resins. A 50 L culture-derived raw material was subjected to polishing purification using this resin after AFF purification, achieving high purity of 91.1% in one purification run and 97.4% in two purification runs. This resin is effective as a scalable empty particle removal technology that can replace gradient ultracentrifugation and contribute to the production of high-purity (approximately 95% full) and high-yield AAV formulations. *This document was presented at BioProcess International 2024, Boston.
Development of a Cellulose Monolith-like Particle for Sulfate Pseudo-affinity Chromatography Targeting Vaccine Purification
We developed and evaluated a chromatography resin based on cellulose-derived monolithic particles (MLP) with continuous pores for the purification of large biopolymers, particularly virus particles used in vaccines. MLP 1000 DexS, modified with dextran sulfate, exhibited high dynamic binding capacity and recovery through affinity, achieving greater adsorption and impurity removal for influenza A (H1N1) than conventional chromatography resins. Its high strength and low pressure drop make it useful for scale-up, contributing to the efficiency of downstream processes in vaccine manufacturing. *This document was presented at BioProcess International 2024, Boston.
Reducing Host Cell Proteins (HCPs) During Monoclonal Antibody Purification Using Multimodal Chromatography (MMC) resin
We developed and evaluated a novel multimodal chromatography (MMC) resin effective for polishing purification after Protein A chromatography. This chromatography resin uses continuous-pore cellulose-derived monolithic particles (MLPs) as a support, with long-chain alkyl groups modified with primary amines. Optimizing the pore size enables powerful reduction of host-derived proteins (HCPs) even at high loads. Using the resin in flow-through mode, we achieved approximately 99% recovery of monoclonal antibodies (mAbs) and reduced HCPs to a minimum of 8 ppm. Because the effect of residence time is minimal, high flow-rate purification contributes to shorter processing times. *This document was presented at BioProcess International 2024, Boston.
Development of cellulose-derived monolithic particles (MLPs) with continuous pores as a next-generation modality for virus particle purification
We have developed cellulose-based monolithic particles (MLP) with continuous pores specifically designed for the purification of large molecules, such as virus particles used in vaccines and gene therapy drug substances. The porous structure with straight pores (mode radius approximately 1.5 µm) allows easy access to the particle interior, achieving high dynamic binding capacity. MLP 1000 DexS, modified with dextran sulfate, demonstrated high recovery and highly purified removal of contaminants in the purification of influenza A and SARS-CoV-2. Furthermore, AEX-modified resins have been shown to outperform existing products in the separation of AAV empty and filled capsids. We will continue to develop these new chromatographic resins as highly effective for next-generation modalities. *This document was presented at BioProcess International 2025, Boston.
The newly developed Cellufine™ Phosphate HC enables efficient purification of T7 RNA polymerase and other large biomolecules.
Cellufine™ Phosphate HC is a novel, high-adsorption chromatography resin with expanded pore volume. It features a unique ligand consisting of phosphate groups modified on cellulose particles. It is suitable for purifying large nucleic acid-binding proteins (over 30 kDa) and can be efficiently purified without the need for a His tag. For T7 RNA polymerase, the dynamic binding capacity is approximately nine times higher than that of existing products. Purification from E. coli lysate consisted of the following steps: 1) reduction of DNA and impurities with Cellufine™ MAX DEAE, 2) affinity purification with Cellufine™ Phosphate HC, and 3) endotoxin reduction with Cellufine™ ET Clean L. These three purification steps rapidly achieved high activity and purity. *This document was presented at BioProcess International 2025, Boston.
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