Ideal for purifying pneumococcal vaccine
Cellufine™ MAX Q-hv
Cellufine™ MAX Butyl HS

Cellufine™MAX Q-hv
Cellufine™MAX Butyl HS

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.
By using two-step chromatography purification to purify the capsular polysaccharide, it is possible to purify the vaccine to a high degree of purity. 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.

Features of Cellufine™ MAX Q-hv

Cellufine MAX Q-hv is a strong anion exchanger. There are several strong anion exchangers in Cellufine's product line, and we will introduce a comparison between them. Cellufine MAX Q-hv has optimized ligand concentration for optimal purification of capsular polysaccharide.


Comparison of Cellufine™ MAX Q series
Grade Cellufine™ MAX Q-r Cellufine™ MAX Q-h Cellufine™ MAX Q-hv
Base resin Highly cross-linked cellulose with dextran scaffold
Particle size ca. 40~130 μm
Lignad Strong anion / -N+(CH3)3
Ion exchange capacity (meq/ml) 0.10~0.20 0.13~0.22 0.04~0.07
DBC BSA (mg/ml) 110 180 120
Flow velocity 600 cm/h(0.3 MPa), I.D.30 cm-L20 cm, pure water at 24 ºC
pH stability pH 2 - 12
Chemical stability Stable all commonly used aqueous buffers
CIP 1 M NaOH 0.5 M NaOH
Storage 20 % ethanol

Features of Cellufine™ MAX Butyl HS

Cellufine™ MAX Butyl HS is a hydrophobic interaction chromatography resin with a butyl group. There are several HIC resin in Cellufine's product line, and we will introduce a comparison between them. Cellufine™ MAX Butyl HS has optimized ligand concentration for optimal purification of capsular polysaccharide.


Comparison of Cellufine™ MAX Butyl series
Grade Cellufine™ MAX Butyl Cellufine™ MAX Butyl HS
Base resin Highly cross-linked cellulose
Particle size ca. 40~130 μm
Ligand Low conc. butyl group High conc. butyl group
BSA adsorption capacity (mg/ml) 9 13
BSA elution efficiency (%) 70 36
Operating pressure <0.3 MPa
pH stability pH 2 - 13
Chemical stability Stable in commonly used buffers
CIP 1 M NaOH
Storage 20 % Ethanol

Protein Separation Performance for Cellufine MAX Butyl HS

Protein Separation Performance for Cellufine MAX Butyl HS
Column
6.6 mm ID x 30 mmL (1.0 ml)
Buffer A
10 mM Na phosphate, 1.5 M ammonium sulfate, pH 7.0
Buffer B
10 mM Na phosphate, pH 7.0
Protein
Ribonuclease A, Cytochrome C, Lysozyme

Flow property of Cellufine™ MAX Butyl HS

Flow property of Cellufine MAX Butyl HS
Column
2.2cm I.D. x 20 cm L
Temperature
24 ± 1 ℃
Mobile phase
Pure water

Purification of capsular polysaccharide of Streptococcus pneumoniae serotype 19F

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. To overcome these disadvantages, a lot of improved processes are proposed. Some of these processes include chromatography processes. 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.

Pneumococcal vaccine purification process
PS
Recovery %
PS Purity % Protein
μg/mL
Nucleic acid
μg/mL
Protein
/ PS %
Nucleic acid
/ PS %
Load sample - 46 73 772 10 106
MAX Butyl HS
after purification
89 54 M.D. 436 0 84
MAX Q-hv
after purification
98 99 N.D. 4 0 1
The HIC step was run in flow-through mode at 0.212 mL/min (residence time 5.0 min). The pool was concentrated and diafiltrated by the cross-flow ultrafiltration device (PES, 100,000 MWCO) with MilliQ water. The AEX step was run in bind and elute mode at 0.424 mL/min (residence time 2.5 min).

Chemicai stability

Stable in the following solvents and autoclaves.

  • Ethanol (70%)
  • Sodium hydroxide (0.5M)
  • Isopropanol (30%)
  • Surfactant
  • Guanidine hydrochloride (6M)
  • Autoclave (121 ºC, 20 min)
  • Urea (6M)

Custom Resin Development Services

JNC offers a custom resin development service for optimization of current products to meet stringent or enhanced performance working directly with large and small customers. In addition, JNC also offers custom resin development services to support new applications of JNC resins.