BG-CHT - 汇融科技

CERAMIC HYDROXYAPATITE

BETAGENE BG-CHT

Specs

CAS号	EC号	MDL编号	货号
9025-65-4	3.1.30.2(BRENDA，IUBMB)	MFCD00131010	GA01.01

DATASHEET

MSDS

COA

BG-CHT by BETAGENE

So intact, yet so affordable

Introduction

Hydroxyapatite, also known as basic calcium phosphate, is the natural mineralization of calcium apatite (Ca₅(PO₄)₃(OH)). But it is often written as Ca₁₀(PO₄)₆(OH)₂ to highlight that it is made up of two parts - Hydroxyl and Apatite. OH-hydroxyl can be replaced by fluoride, chloride and carbonate ions to form fluoro- or chloro-apatite.

Flow rate and scale of crystalline hydroxyapatite in industrial production are limited due to the stiffness of crystal structure,. In the 1980s, scientists created stronger porous hydroxyapatite materials by burning them at high temperatures. Since then hydroxyapatite materials can withstand higher flow rates and pressures in large-scale industrial production, greatly improving production efficiency. This type of high-temperature-processed hydroxyapatite material is commonly known as ceramic hydroxyapatite. Now ceramic hydroxyapatite has been widely used in the purification and preparation of biological macromolecules, the adsorption and removal of heavy metal ions and bone repair materials.

Mechanism

Hydroxyapatite consists of two binding sites: Ca²⁺and PO₄^3-. These binding sites are regularly distributed throughout the crystal structure. PO₄^3- ions are bonded with positively charged proteins through cation exchange. Ca²⁺ ions are metal chelated to the free carboxyl clusters of negatively charged proteins.

Cation exchange: Positively charged protein amino groups can interact ionically with PO₄^3- and repel Ca²⁺. Ion exchange interactions are affected by the concentration of neutral salts (such as NaCl) in the eluting buffer or by the concentration of phosphate in the equilibrium solution. Interactions will weaken with the increase of pH. Therefore, increasing the concentration of salt and phosphate or increasing pH will weaken the interaction between the target protein and PO₄^3-.

Calcium-metal chelation: Ca²⁺ can metal-chelate with carboxyl clusters or phosphoryl groups on the surface of proteins or other molecules (such as nucleic acids) while simultaneously repelling PO₄^3-. The binding force is 15-60 times that of normal ion exchange. Therefore, this binding mode is less affected by ions and is not sensitive to NaCl. This binding force increases the metal chelating ability of calcium affinity with the increase of ionic strength, because ionic strength shields the mutual repulsion between functional groups on molecular surface and PO₄^3-. The metal affinity interactions can be eluted by sodium phosphate.

Features

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BG-CHT is a ceramic hydroxyapatite developed specially for purification of bio-macromolecules. BG-CHT has extraordinary batch stability and purification-binding characteristics, which is produced by carefully selected raw materials. Our BG-CHT material has the advantages of spherical particles, strong mechanical properties, large specific surface area, stable binding assay and mass production. It has the potential to substitute for existing domestic and foreign hydroxyapatite materials.

Type

We provide Type I and Type II BG-CHT in two particle sizes (20 μm and 40 μm). Type I CHT is homogeneous spherical particles fully sintered at 400 ℃, and Type II CHT is homogeneous spherical particles fully sintered at 700 ℃. Both types are hydroxyapatite in nature. The binding and elution characteristics of both types are similar to those of crystalline hydroxyapatite, but there are some differences: Type I has a higher protein binding capacity and is more sensitive to acidic proteins. Type II has a lower capacity and is more sensitive to eluting proteins and nucleic acids. Due to its low affinity for albumin, Type II is generally more suitable for purifying various immunoglobulins.

40μm

20μm

Application

BG-CHT is an ultrapure 10nm × 100nm hexagonal cross-section crystal developed and produced by BETAGENE Genetic with top quality materials through proprietary technology. The crystal is condensed into particles, which are then heated to form stable connections at the crystal contact points. The particle size and porosity are strictly controlled to make the particles easy to pack, and finally to achieve excellent separation performance.

The hybrid purification mechanism provides unique selectivity for the separation of various biomolecules of similar properties that cannot be separated by other mediums. BG-CHT can be used for the separation of host proteins (HCP), shed Protein A, antibody dimers and polymers, nucleic acids and viruses, and can be used in each purification stage of capture, intermediate purification or purification.

Regenerate

Regeneration

Washing the CV with 500 mM neutral phosphate or potassium phosphate solution or 400 mM trisodium phosphate (pH 11-12) for 3-5 CV. The column can also be washed with neutral 1-2 M KCl, NaCl, 6M urea or 8M guanidine HCl containing 5 mM phosphate.

Storage

Unused BG-CHT should be stored at room temperature, sealed tight. Once wetted, store in 0.1 M NaOH at room temperature. If using less than 100 mM NaOH, add 10 mM phosphate. Used CHT, after regeneration and sterilization, should be stored in 1 M or less NaOH in shades at room temperature.

Sanitization

The column can be sanitized in up to 2M NaOH, and also could be stored in 0.1-1M NaOH.

QA

Quality Assurance

Items	Specifications	Standard
Appearence	White free flow powder	Visual
Lysozyme binding assay	≥ 12.5 mg/g power	Enterprise standard
Protein sample separation	Passes test
Particle size (mean)	30 - 50 μm
Tapped density	0.6 - 0.7 g/mL
FTIR	Passes test
Endotoxin	≤ 0.01 EU/kU

Appearence：White free flow powder

Lysozyme binding assay ≥ 12.5mg/g

Protein sample separation：Passes

Particle sizes (mean)：30-50 μM

Endotoxin：≤ 0.01 EU/kU

Tapped density：0.6 - 0.7 g/mL

FTIR: Passes

According to enterprise standard test

安全数据

储存分类代码	WGK	闪电（F)
10-Combustible liquids	WGK 1	Not Applicable
闪电（C)	个人防护装备
Not Applicable	Eye shields, gloves

Specs

COMPOSITION

HYDROXYAPATITE

PARTICLE SIZE（MEAN）

20/40/80 μM；MEAN：43 μM

CAT.No

GC01.01/GC01.02

APPEARENCE

WHITE FREE FLOWING POWDER

ENDOTOXINES

< 0.01 EU/mg

FUNCTIONAL GROUPS

Ca⁺，PO^-，OH

pH STABILITY

6.5 - 14

BASE STABILITY

> 1 YEAR IN 1 N NaOH

TAPPED DENSITY

0.63 g/mL

LYSOZYME BINDING ASSAY

14 mg/g

DYNAMIC BINDING CAPACITY

TYPE I：25 - 60 mg POLYCLONAL IgG/mL CHT；TYPE II：15 - 25 mg POLYCLONAL IgG/mL CHT

REGENERATION

0.4 M SODIUM PHOSPHATE，pH 7-7.5；1 M TRISODIUM PHOSPHATE，pH 11-12

AUTOCLAVABILITY（BULK）

121°C，20 MIN IN PHOSPHATE BUFFER，pH 7

TYPICAL LINEAR FLOW RATE

500-1000 CM/HR

比活性高
解决核酸彻底
适应面广
耐受多种实验条件

Specs
FAQs

Composition

Ceramic hydroxyapatite

Cat. No.

GC01.01，GC01.02

White Free Flowing Powder

Physical Appearance

Lysozyme binding assay

14 mg/g power

Particle size(mean)

20、40、80μm；Mean：43μm

Functional groups

Dynamic binding capacity

Type I：25 - 60 mg polyclonal lgG/mL CHT；Type II：15 - 25mg policlonal IgG/mL CHT

pH stability

6.5 - 14

Tapped density

0.63 g/ml

Regeneration

0.4 M sodium phosphate，pH 7-7.5；1 M trisodium phosphate，pH 11-12

At least 1 year in 1 N NaOH

Base stability

Autoclavability (bulk)

121°C，20 min in phosphate buffer，pH 7

Endotoxines

< 0.01 EU/kU

Typical linear flow rate range

Ca⁺，PO^-，OH^-

50-1000 cm/hr

What are the main features of BG-CHT?

BG-CHT is a ceramic hydroxyapatite developed specially for purification of bio-macromolecules. BG-CHT has extraordinary batch stability and purification-binding characteristics, which is produced by carefully selected raw materials. Our BG-CHT material has the advantages of spherical particles, strong mechanical properties, large specific surface area, stable binding assay and mass production. It has the potential to substitute for existing domestic and foreign hydroxyapatite materials.

How to configure the buffer?

Most buffers should contain at least 5 mM phosphate. Basic proteins can be eluted with a step or linear gradient using pH 7.2 phosphate buffer with 1 M NaCl. Acidic proteins and nucleic acids are eluted with a step or linear gradient using phosphate. It is critical that the pH of both the buffers and the efﬂuent are at pH 6.5 or higher.

How to prolong BG-CHT column life?

All buffers, including the storage solution, should contain 5 Mm phosphate at a pH ≥ 6.5. High pH buffers should be used for all applications. Adding calcium ions at ppm level can improve the firmness of the packing. When first eluting with NaCl, the pH will decrease due to hydrogen ion shedding, so adding a co-buffer, such as MES, can improve the firmness of the packing. Regenerating with 500 mM phosphate removes adsorbed protein and can be stored in 0.1 M NaOH. If the NaOH concentration is less than 0.1 M, add 5 mM phosphate. Preserving the column with NaOH prevents carbonate precipitation on the column surface. Avoiding abrupt changes in flow rate reduces pressure shock.

What is expected HETP and asymmetry of BG-CHT?

When equilibrated with phosphate buffered saline and tested with a 2.5% test of 1 M NaCl in phosphate buffer, the typical HETP values for columns with a diameter greater than 5 cm for 40 μm BG-CHT range from 0.016–0.021 cm, and the range for 80 μm is 0.032–0.041 cm. Asymmetry values from 0.8–2.3 are usually acceptable.

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