CHT™ Ceramic Hydroxyapatite Type II Media

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Overview

Specifications
  Type I Type II
Functional groups Ca2+, PO4, OH Ca2+, PO4, OH
Observed dynamic binding capacity lysozyme (Lys) ≥25 mg Lys/g CHT ≥12.5 mg Lys/g CHT
Nominal pore diameter 600–800 A 800–1,000 A
Maximum backpressure 100 bar (1,500 psi) 100 bar (1,500 psi)
Nominal mean particle size 20 ± 2, 40 ± 4, and 80 ± 8 µm 20 ± 2, 40 ± 4, and 80 ± 8 µm
Bulk density 0.63 g/ml 0.63 g/ml
Ceramic Hydroxyapatite (CHT™) for Commercial Purification

Ceramic hydroxyapatite is one of the leading media for biomolecule purification. It's a spherical ceramic form of crystalline hydroxyapatite and this form allows it to be used in production-scale columns at high flow rates while maintaining its unique separation properties.

CHT Binding Mechanism

CHT Ceramic Hydroxyapatite Mechanism

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 Ceramic Hydroxyapatite
  Type I Type II
Functional groups Ca2+, (PO4)3- Ca2+, (PO4)3-
Dynamic binding capacity >25 mg lysozyme per gram CHT >12.5 mg lysozyme per gram CHT
Typical IgG binding capacities at 500 cm/hr 25–60 mg/ml 15–25 mg/ml
Particle sizes available 20, 40, and 80 µm (nominal) 20, 40, and 80 µm (nominal)
Nominal pore diameter 600–800 Å 800–1,000 Å
Maximum operating pressure 100 bar (1500 psi) 100 bar (1500 psi)
Recommended operating flow rates 10–1,500 cm/hr 10–1,500 cm/hr
pH stability 6.5–14 6.5–14
Regeneration 0.4-1.0 M phosphate buffer
0.5 M sodium phosphate pH 7
1.0 M trisodium phosphate pH 11–12
0.4-1.0 M phosphate buffer
0.5 M sodium phosphate pH 7
1.0 M trisodium phosphate pH 11–12
Sanitization in 1–2 N NaOH in 1–2 N NaOH
Autoclavability 20 min at 121°C 20 min at 121°C
IgG Protocol Using CHT

Purification of IgG on UNOsphere Rapid S 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

Purification of IgG using CHT Ceramic Hydroxyapatite. Read more.


Globular Protein Protocol Using CHT
Protocol II: Globular Proteins

Protocol II: Globular Proteins

Flow rate: 300 cm/hr
Buffer A: 5 mM NaPO4, 150 mM NaCl, pH 6.8
Buffer B: 500 mM NaPO4, 150 mM NaCl, pH 6.8
Buffer C: 500 mM NaPO4, pH 6.8

Equilibrate the column with approximately 10 column volumes of buffer A. Prepare the sample, adjusting the pH and conductivity to those of buffer A. Load the sample in 5 mM NaPO4, pH 6.8; wash the column with approximately 5 column volumes of buffer A; and elute with approximately 20 column volumes of buffer B linear gradient. Slope and amplitude can be adjusted based on initial results. After elution clean the column with approximately 5 column volumes of buffer C followed by a sanitation step using approximately 5 column volumes of 1 M NaOH.

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
Protocol III Plasmids

3.2.3 Protocol III: Plasmids

Flow rate: 300 cm/hr

Buffer A: 10 mM NaPO4, 1 mM EDTA, pH 7.0

Buffer B: 400 mM NaPO4, 1 mM EDTA, pH 7.0

Equilibrate the column with approximately 10 column volumes of buffer A. Prepare the sample, adjusting the pH and conductivity to those of buffer A. Load the sample in 0.5 M NaCl pH 7.0; wash the column with approximately 5 column volumes of buffer A; and elute with approximately 20 column volumes of buffer B linear gradient. Slope and amplitude can be adjusted based on initial results. After elution clean the column with approximately 5 column volumes of buffer A followed by a sanitation step using approximately 5 column volumes of 1 M NaOH.

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

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. This 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 dimers 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

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

For more information, request bulletin 5709.

Regulatory support files are available upon request. Bio-Rad Laboratories is an ISO 9001-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:   $155.00
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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

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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:   $981.00
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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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Foresight™ CHT™ Type II Plates

732-4718
2 x 96-well plates, 20 μl, filter plate prepacked with CHT ceramic hydroxyapatite Type II multimodal chromatography media, 40 μm particle size

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Foresight™ CHT™ Type II RoboColumn Unit

732-4825
1 row of 8 columns, 200 μl, RoboColumn unit prepacked with CHT ceramic hydroxyapatite Type II multimodal chromatography media, 40 μm particle size

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Foresight™ CHT™ Type II RoboColumn Unit

732-4826
1 row of 8 columns, 600 μl, RoboColumn unit prepacked with CHT ceramic hydroxyapatite Type II multimodal chromatography media, 40 μm particle size

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Foresight™ CHT™ Type II Column

732-4736
Pkg of 1, 1 ml, column prepacked with CHT ceramic hydroxyapatite Type II multimodal chromatography media, 40 μm particle size

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Foresight™ CHT™ Type II Column

732-4756
Pkg of 1, 5 ml, column prepacked with CHT ceramic hydroxyapatite Type II multimodal chromatography media, 40 μm particle size

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Number Description Options
6549 Mammalian Virus Purification Using Ceramic Hydroxyapatite, Rev A Click to download [ Add to Cart (Free) ]
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) ]
5709 How CHT Ceramic Hydroxyapatite Works, 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) ]
2156 CHT Ceramic Hydroxyapatite — A New Dimension in Chromatography of Biological Molecules, Rev C Click to download [ Add to Cart (Free) ]
5739 Packing and Slurry-in-Place Procedures for CHT Ceramic Hydroxyapatite Using a Bio-Rad InPlace Process-Scale Chromatography Column, Rev A Click to download [ Add to Cart (Free) ]
2731 Plasmid Purification Using CHT Ceramic Hydroxyapatite Support, Rev A Click to download
2780 Purification of Murine IgG1 Using UNOsphere S and CHT Ceramic Hydroxyapatite Chromatography, Rev A 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
1986 Effect of pH on Gradient Elution of Proteins on Two Types of CHT Ceramic Hydroxyapatite, Rev B Click to download [ Add to Cart (Free) ]
1985 Clearance of Murine Leukemia Virus From a Chimeric Monoclonal Antibody Using Ion Exchange Chromatography, Rev B. Click to download [ Add to Cart (Free) ]
2940 Removal of Aggregate From an IgG4 Product Using CHT Ceramic Hydroxyapatite, Rev A Click to download
2849 Protein A Removal From IgG on CHT Ceramic Hydroxyapatite Support, Rev A Click to download
2535 Purification of Recombinant Human alpha 1-Antitrypsin From Transgenic Milk Using CHT Ceramic Hydroxyapatite, Rev B Click to download
2575 Purification of a Calcium-Binding Protein Using CHT Ceramic Hydroxyapatite Click to download
2946 Plasmid DNA Adsorption Behavior on CHT Ceramic Hydroxyapatite Type II Support, Rev A Click to download
5662 Process Chromatography Applications Laboratory Brochure, Rev B Click to download
5584 Bio-Scale Mini Cartridges, Product Information Sheet, Rev B Click to download