LC pH离子淌度试剂: 色谱专用

pH Gradient Buffer for LC

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LC pH离子淌度试剂是一种有机盐线性PH值梯度离子交换洗脱试剂。 适用于mAb抗体， 糖蛋白, PTM修饰蛋白等通过离子交换色谱分离或纯化的通用流动相。在基于pH离子淌度试剂的离子交换色谱中，起始缓冲液的 pH 值保持在恒定水平，以确保带电荷蛋白质与固定相相反的电荷离子与其结合。通过线性改变缓冲液pH 值来洗脱蛋白质。可以快速分离蛋白药物的酸性碱性峰并表征蛋白药物Native 一级和亚型结构。一种通用的 mAb分离纯化新技术。

产品用途：

• 生物治疗蛋白电荷异质性分析

• 预测出带电荷异构体的 pH 和预期保留情况

• Native亚型结构的分离和表征

• Native整体蛋白的纯化

• 分离和纯化蛋白

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产品特点：

• 质谱级干粉试剂

• pH 洗脱线性好

• 梯度稳健且可重现

• 较窄的 pH 范围实现更高分辨率的分离

Part No.                          Name                                         Size

HLS PH01LCA              LC pH Gradient Buffer A.     6.1 g

HLS PH01LCB              LC pH Gradient Buffer B.     6.65 g

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Description: pH gradient buffer is an innovative elution buffer (or mobile phase) that changes pH linearly for separation and purification of mAb antibodies, glycoproteins and PTM modified proteins by ion exchange (IEX) chromatography. In pH gradient buffer-based IEX chromatography, the pH of the initial buffer is kept at a constant level to ensure that the charged proteins bind to oppositely charged ions of the stationary phase. Then, the protein is eluted by linearly changing the pH of the buffer so that the protein has a zero net charge and can elute from the column.

Physical Appearance: Lyophilized powder

Reagent Composition: Each bottle contains 2.5 mg BT Surfactant, 850 mg NaCl, and 6 g sulfonate.

Resuspension: Dissolve reagents in 500 mL deionized, double-distilled water.

pH Adjustment: At room temperature, use pH meter to ensure the pH error is less than 0.2, and use 2 M NaOH or hydrochloric acid to adjust the pH.

Elution Buffer (mAb): pH Buffer A, pH 5.5

Elution Buffer B (maB): pH Buffer B, pH 10.2

Storage Conditions: In 4 °C refrigerator as lyophilized powder.

Shelf Life: 24 months at 4 °C as powder; 7 days at 4 °C as solution.

Linear pH Range: Linear correlation coefficient > 0.99 at pH 5.5 - 10.2 in columns of IEX chromatography.

Reference Procedure ofr mAb subunit and intact protein:

Column: SCX, 10 μm, 2.1 * 250 mm or 2.1 * 50 mm                  Flow rate: 0.3 mL/min

Eluent A: 1X mAb pH buffer A (pH 5.5)                                    Eluent B: 1X maB pH Buffer B (pH 10.2) Gradient: 0 -1 min, 0% B; 1 - 31 min, 0 - 100% B;                                                                      31 - 34 min, 100% B; 34 -40 min, 0%B Temperature: 30 °C FLR detection: Ex: 280 nm, Em: 360 nm

UV detection: 280 nm                                                              Load: 0.2 - 10 μg

Inject volume: 4 μL

| Quality Control

Purity: In ion mobility buffer, the BT Surfactant is mass spectrometry reagents, and other components are pure chromatography reagents with < 0.01 absorbance at 280 nm, analyzed by liquid chromatography.

pH Accuracy: pH 5.5 for mAb pH Buffer A and pH 10.2 for maB pH Buffer B with error less than 0.2.

pH Elution Linearity: The linear correlation coefficient of 227 nm absorbance > 0.999 with zero volume injection and SCX ion exchange chromatography system in 30 min run time at 30 °C column temperature.

pH Elution Degree of Seperation: Linear elution degree of separation > 1 with 5 pKa protein mAb: IGg A1, Ribonulease A, Ribonuclease B, and Cytochrome C.

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