CHT Ceramic Hydroxyapatite Type II Support

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Overview

Specifications
Functional groups Ca2+, PO43-, OH
Particle sizes available 20, 40 and 80 µm (nominal)
Recommended linear flow rate 50–1,000 cm/hr
Operating pH range 6.5–14
Chemical compatibility (>24 hr) 1 M NaOH, 8 M urea, 8 M guanidine-HCl, ethanol, methanol, 100% acetonitrile
Regeneration 0.4–1.0 M phosphate buffer, 0.5 M sodium phosphate pH 7
1.0 M trisodium phosphate pH 11–12
Sanitization 1–2 N NaOH
Autoclavability (121µC, 20 min) Yes
Packing density (g/ml packed bed) 0.63 g/ml
  Type I Type II
Dynamic binding capacity > 25 mg lysozyme/g > 12.5 mg lysozyme/g
Typical IgG binding capacities at 500 cm/hr 25–60 mg/ml 15–25 mg/ml
Nominal pore diameter 600–800 Å 800–1,000 Å
Maximum operating pressure 100 bar (1,500 psi) 100 bar (1,500 psi)
CHT Binding Mechanisms

Hydroxyapatite contains two types of binding sites, positively charged calcium and negatively charged phosphate groups. These sites are distributed regularly throughout the crystal structure of the matrix. Solute species dominantly interact through cation exchange via the phosphate groups and/or metal affinity via the calcium atoms.

Characteristics of Cermaic Hydroxyapatite
  Type I Type II
Observed dynamic binding capacity IgG 25–60 mg IgG/ml CHT* 15–25 mg IgG/ml CHT**
Typical linear flow rate range 50–1,000 cm/hr  
pH stability*** 6.5–14 pH  
Base stability at least 21 months in 1 N NaOH  
Regeneration 500 mM sodium phosphate, pH 7  
  1,000 mM trisodium phosphate, pH 11–12  
Autoclavability (bulk) 121°C, 20 min in phosphate buffer, pH 7  
Sanitization 1–2 N NaOH  
Recommended column storage 0.1 M NaOH  
Shelf life (dry, unused material) 85 months stored dry, sealed and at room temperature  

* 40 µm particles, 300 cm/hr, 5 mM sodium phosphate, pH 6.5
** 40 µm particles, 300 cm/hr, 5 mM sodium phosphate, pH 6.5
*** For pH 5.5–6.0, see Section 3.1.1 Buffers and Table 1

IgG Protocol Using CHT

Optimizing the bioprocess requires management of multiple variables ranging from matrix interaction, elution characteristics, scale-up from bench, regulation and drug safety requirements, and process robustness and economics. This IgG protocol has been developed as general starting guidelines for the purification of most proteins and nucleic acids and may help to reduce time spent in methodology development.

Purification of IgG using CHT

Elution of monoclonal antibody in a 40 column volume linear gradient to 1 M NaCl.

Globular Protein Protocol Using CHT

Optimizing the bioprocess requires management of multiple variables ranging from matrix interaction, elution characteristics, scale-up from bench, regulation and drug safety requirements, and process robustness and economics. This protocol has been developed as general starting guidelines for the purification of most proteins and nucleic acids and may help to reduce time spent in methodology development.

Plasmid Protocol Using CHT

Optimizing the bioprocess requires management of multiple variables ranging from matrix interaction, elution characteristics, scale-up from bench, regulation and drug safety requirements, and process robustness and economics. This protocol has been developed as general starting guidelines for the purification of most proteins and nucleic acids and may help to reduce time spent in methodology development.

Acidic Proteins Protocol Using CHT

Optimizing the bioprocess requires management of multiple variables ranging from matrix interaction, elution characteristics, scale-up from bench, regulation and drug safety requirements, and process robustness and economics. This protocol has been developed as general starting guidelines for the purification of most proteins and nucleic acids and may help to reduce time spent in methodology development.

CHT ceramic hydroxyapatite, (Ca5(PO4)3OH)2, support is a leading purification medium for today's demanding downstream process industry. Its mixed-mode support offers unique selectivities and often separates biomolecules that appear homogeneous using other chromatographic methods. The diverse binding capabilities of CHT for host cell proteins, leached protein A, antibody dimmers and aggregates, nucleic acids, and viruses allow its use at any stage from initial capture to final polishing.

The robust properties of CHT ceramic hydroxyapatite improve efficiency, yield, and financial value through:

  • Excellent capture at elevated flow rates, enabling processing at all scales
  • Large capacity for high-titer upstream feedstocks
  • Exceptional selectivity, allowing a two-step chromatographic process

Hydroxyapatite (Ca5(PO4)3OH)2 is a form of calcium phosphate used in the chromatographic separation of biomolecules. Sets of five calcium doublets (C-sites) and pairs of hydroxyl-containing phosphate triplets (P-sites) are arranged in a repeating geometric pattern. Repeating hexagonal structures can be seen in electron micrographs of the material; space-filling models and repeat structure from Raman spectroscopy have also been constructed.

Two types of CHT ceramic hydroxyapatite, Type I and Type II, are available in three particle sizes, 20, 40, and 80 µm. Both types have elution characteristics similar to crystalline hydroxyapatite, but also have some important differences:

  • Type I, sintered at 400°C, has a higher protein binding capacity and better capacity for acidic proteins
  • Type II, sintered at 700°C, has a lower protein binding capacity, but offers better resolution of nucleic acids and certain proteins; it also has a very low affinity for albumin and is particularly well-suited for the purification of many species and classes of immunoglobulins

Regulatory support files are available upon request. Bio-Rad Laboratories is an ISO9001 registered corporation.

CHT Ceramic Hydroxyapatite #158-2200
CHT Ceramic Hydroxyapatite

158-2200
10 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 20 μm particle size

List Price:   $123.00
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CHT Ceramic Hydroxyapatite #157-2000
CHT Ceramic Hydroxyapatite

157-2000
100 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 20 μm particle size

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CHT Ceramic Hydroxyapatite #157-2100
CHT Ceramic Hydroxyapatite

157-2100
1 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 20 μm particle size

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CHT Ceramic Hydroxyapatite #157-2500
CHT Ceramic Hydroxyapatite

157-2500
5 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 20 μm particle size

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CHT Ceramic Hydroxyapatite #158-4200
CHT Ceramic Hydroxyapatite

158-4200
10 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 40 μm particle size

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CHT Ceramic Hydroxyapatite #157-4000
CHT Ceramic Hydroxyapatite

157-4000
100 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 40 μm particle size

List Price:   $871.00
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CHT Ceramic Hydroxyapatite #157-4100
CHT Ceramic Hydroxyapatite

157-4100
1 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 40 μm particle size

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CHT Ceramic Hydroxyapatite #157-4500
CHT Ceramic Hydroxyapatite

157-4500
5 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 40 μm particle size

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CHT Ceramic Hydroxyapatite

158-8200
10 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 80 μm particle size

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CHT Ceramic Hydroxyapatite

157-8000
100 g, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 80 μm particle size

List Price:   $784.00
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CHT Ceramic Hydroxyapatite #157-8100
CHT Ceramic Hydroxyapatite

157-8100
1 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 80 μm particle size

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CHT Ceramic Hydroxyapatite #157-8500
CHT Ceramic Hydroxyapatite

157-8500
5 kg, CHT ceramic hydroxyapatite multimodal chromatography media, Type II, 80 μm particle size

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Number Description Options
5739 Packing and Slurry-in-Place Procedures for CHT Ceramic Hydroxyapatite Using a Bio-Rad InPlace Process-Scale Chromatography Column, Rev B Click to download
6278 High-Throughput Calcium Analysis of Hydroxyapatite Column Effluent, Rev A Click to download [ Add to Cart (Free) ]
6086 Ceramic Hydroxyapatite Application Guide for Process Development and Scale-Up, Rev C Click to download [ Add to Cart (Free) ]
6067 EDTA Complexometric Titration of Hydroxyapatite Column Effluent, Rev B Click to download
RP0033 A Ceramic Hydroxyapatite–Based Purification Platform — Simultaneous Removal of Leached Protein A, Aggregates, DNA, and Endotoxins from MAbs Click to download [ Add to Cart (Free) ]
2524 Purification of Horse IgGT Using Macro-Prep DEAE and CHT Ceramic Hydroxyapatite Type I Supports, Rev C Click to download
2774 Purification of Transgenic Antibody From Corn Seed Using UNOsphere S and CHT Ceramic Hydroxyapatite Supports, Rev A Click to download
2026 Evaluation of Different Approaches for the Chromatographic Purification of Monoclonal Antibodies Click to download
5853 Chimeric IgG Monoclonal Antibody Purification: A Comparative Study Using CHT Ceramic Hydroxyapatite and CFT Ceramic Fluoroapatite Chromatographic Media, Rev A Click to download [ Add to Cart (Free) ]
5667 CHT Ceramic Hydroxyapatite Product Information Sheet, Rev B Click to download [ Add to Cart (Free) ]