pH离子淌度试剂是一种创新的线性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，使蛋白质转变为净零电荷并从柱中洗脱。
试剂成分: 每瓶含2.5mg BT Surfactant; 850mg NaCl; 和6g 磺酸盐
试剂溶解：加500ml超纯去离子水溶解。
pH值确认：在室温条件下，使用pH酸度计确认pH值误差<0.2;必要时使用2M NaOH 或盐酸微调节
洗脱液(mAb): pH Buffer A, pH=5.5; 洗脱液B(mAb): pH Buffer B, pH=10.2
pH 线性范围: 在离子交换色谱柱（IEX)中，pH 5.5 to pH 10.2线性相关系数大于0.99
推荐的mAb subunit or Intact Protein离子交换色谱条件:
Column: SCX, 10μm, 2.1 *250 mm or 2.1*50mm
Flow rate: 0.3mL/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%
Temperature: 30 ℃
UV detection: 280 nm
Inject volume: 4 μL
FLR detection: Ex: 280,nm Em: 360nm
Load: 0.2-10ug
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.
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
Linear pH Range: Linear correlation coefficient > 0.99 at pH 5.5 - 10.2 in columns of IEX chromatography.
Reference Procedure of 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: 1×mAb pH buffer A (pH 5.5)
Eluent B: 1× 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
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