MS 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，可以实现IEX-MS联用系统分析。该离子对试剂在质谱电离时会自动降解，不会有离子抑制现象。
试剂成分: 每瓶含0.3-1g 醋酸铵; 0.2g 有机酸
试剂溶解：加100ml超纯去离子水溶解。
mAb pH离子淌度试剂：用色谱级浓氨水调节 MS pH试剂A: pH=5.5； MS pH试剂B: pH=8.5
洗脱液A (mAb): pH Buffer A, pH=5.5; 洗脱液B(mAb): pH Buffer B, pH=8.5
pH 线性范围: 在离子交换色谱柱（IEX)中，pH 5.5 to pH 8.5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: 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
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. This buffer is suitable for IEX-MS to characterize intact mAb or subunit.
Reagent Composition: Each bottle contains 0.3-1 g Ammonium acetate, and 0.2 g Organic acid.
Resuspension: Dissolve reagents in 100 mL deionized, double-distilled water.
pH Adjustment: At room temperature, slowly add LC grade ammonia adjust pH value to ensure the Buffer A pH=5.5; the Buffer B=8.5 for ready mobile phase of LC.
Elution Buffer (mAb): pH Buffer A, pH 5.5
Elution Buffer B (mAb): pH Buffer B, pH 8.5
Linear pH Range: Linear correlation coefficient > 0.99 at pH 5.5 - 8.5 in columns of IEX chromatography.
Reference Procedure for 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 8.5)
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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