Description
This product is composed of a novel high-rigidity agarose matrix coupled with recombinant Protein A (rProtein A) ligand. The rProtein A is expressed and purified from E. coli, ensuring an animal-origin-free production process. The newly designed rProtein A exhibits improved alkali resistance and stronger specific binding to the Fc region of IgG compared to native Protein A. The high-rigidity agarose matrix offers excellent chemical stability and biocompatibility. Through site-specific coupling technology, rProtein A is immobilized onto the matrix, resulting in high binding capacity, high flow rate, superior physicochemical stability, low non-specific adsorption, minimal ligand leakage, and extended service life.This resin is suitable for one-step purification of high-purity antibodies from samples such as ascites, serum, and cell culture supernatant, making it an ideal choice for large-scale antibody production.Aladdin Alkali-tolerant rProtein A Agarose Resin is stored in 20% ethanol, with a settled gel to storage solution ratio of 1:1. The product specification refers to the actual volume of the settled gel.ParameterValueMatrixHigh-rigidity agaroseLigandAlkali-resistant rProtein AAverage Particle Size70 µmDynamic Binding Capacity (6 min residence time)65–80 mg human IgG/mL resinMaximum Flow Rate (25°C)300 cm/hLigand LeakageMaximum Pressure Tolerance0.3 MPa (3 bar)Cleaning-in-Place (CIP)0.1~0.5 M NaOHpH 2~14Storage Conditions2–8°C in 20% ethanol or 2% benzyl alcoholProtocolDue to varying affinity of the resin for antibodies from different species and subtypes, and differences in buffer conditions required to maintain antibody bioactivity, method screening is recommended.A reference purification procedure is provided below:1.Equilibration: Equilibrate the column with 5–10 column volumes (CV) of Buffer A (20 mM phosphate buffer + 0.15 M NaCl, pH 7.0; or 0.05 M borate, 4.0 M NaCl, pH 9.0) until the baseline stabilizes.2.Loading: Load the sample. For solid samples, dissolve in Buffer A. For liquid samples, dialyze against Buffer A prior to loading.3.Washing: Wash the column with 5 CV of Buffer A until the UV baseline returns to baseline levels.4.Elution: Elute bound antibodies using a linear gradient over 10 CV to 100% Buffer B (20 mM citrate, pH 3.0; or 0.1 M glycine, pH 3.0; or 20 mM acetate, pH 3.0).5.Neutralization: Collect the elution fractions and immediately neutralize using a neutralization buffer (e.g., 1.0 M Tris-HCl, pH 9.0, or other suitable buffer) to bring the antibody solution to a stable pH.6.Regeneration & Cleaning-in-Place (CIP): After multiple cycles, regenerate the column using one of the following methods:Option 1: Wash with 3–5 CV of 0.1 M acetic acid or 0.1 M acetic acid/20% ethanol. Rinse with buffer until neutral pH is reached before reuse.Option 2: Wash with 3–5 CV of 0.1–0.5 M NaOH. Rinse with 3–10 CV of purified water. Equilibrate with buffer to neutral pH before reuse