In the biopharmaceutical industry, in order to ensure high quality of products, it is often necessary to carry out clean-in-place (CIP) of chromatography systems. While the column efficiency and transfer rate of the column are also reduced due to the irreversible precipitation of denatured proteins, lipids, and DNA molecules in the sample.
0.1-1 N NaOH can effectively solve the above problems. In addition to sterilizing and inactivating viruses, it can also dissolve denatured proteins, lipids, and DNA molecules deposited on the top of the column, to extend the service life chromatography media. However, natural or recombinant protein A ligands are not tolerant to NaOH, easily getting denatured or shed.
Alkaline-tolerant Protein A Affinity Chromatography Media (Monofinity A Resin, Cat. No. L00433) is alkali-resistant Protein A affinity purification media developed by Genscript to meet the market needs. The product is made of specially engineered alkali-resistant Protein A ligand cross-linked to a novel agarose gel matrix. It has high dynamic binding capacity and low level of ligand shedding, which is suitable for purification of industrial-grade monoclonal antibodies and therapeutic Fc-Fusion proteins.
Specification
Product Name |
Monofinity A Resin |
Composition |
Highly cross-linked 4% agarose |
Average Particle Size |
90μm |
Ligand |
Alkali-tolerant recombinant protein A (E. coli) |
Dynamic Binding
Capacity (DBC) |
30 mg human lgG/ ml
(4 min RT with sample hIgG concentration at 5 mg/ ml) |
Ligand Leakage |
<10 ng/ mg |
Pressure/Flow Specification |
0.3 MPa/ ~1200 cm/ h |
Recommended Flow Rate |
50 to 300 cm/ h |
Chemical Stability |
Stable in all commonly used buffers |
pH Working Range |
3-12 |
CIP Stability |
0.1-0.5M NaOH |
Storage |
20% Ethanol, 2-8 ℃ |
Example
Lane M: Protein Standard
Lane 1: Purified antibody under reducing conditions
Lane 2: Purified antibody under non-reducing conditions
Figure 1. Electrophoresis of humanized IgG purified from cell culture supernatant by Monofinity A Resin
Dynamic Binding Capacity Performance
Figure 2. Dynamic binding capacity (DBC) versus sample concentration
Figure 3. Dynamic binding capacity (DBC) versus residence time
Figure 4. Breakthrough curve with 5mg/ ml hIgG
Protein A Leaching
Figure 5. Protein A leakage measured at different cycles of CIP (4 batches)
Stability & Storage
Figure 6. Dynamic binding capacity stability of Monofinity A Resin for 200 cycles of CIP (0.1M-0.5 M NaOH)
Figure 7. Shelf life of Monofinity A Resin under different storage conditions