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AbCap A 4FF
Smart-Lifesciences rProtein A Beads 4FF is an affinity chromatography medium designed for the purification of monoclonal antibody, polyclonal antibody and Fc tag fusion protein.The recombinant protein A is coupled to highly cross-linked 4% sepharose gel. This coupling method can improve the binding capacity for immunoglobulin. The static binding capacity of rProtein A Beads 4FF is greater than 40 mg human IgG/ml settled beads. The dynamic binding capacity will vary depending on several factors such as target antibody,flow rate etc.
· Product Description
Table 1. Characteristics of rProtein A Beads 4FF
| Item | Description |
|---|---|
| Matrix Spherical | Highly cross-linked 4% agarose beads |
| Ligand | recombinant protein A |
| Static Binding Capacity | >40mg Rabbit IgG/ml medium |
| Particle size | 45-165um |
| Maximum Pressure | 0.3MPa, 3bar |
| pH | 3-10 |
| Storage Solution | 1×PBS containing 20% ethanol |
| Storage Temperature | 2℃-8℃ |
Table 2. Relative binding strengths of antibodies from various species to protein G and protein A as measured in a competitive ELISA test.
| Species | Subclass | Protein A binding | Protein G binding |
|---|---|---|---|
| Human | IgA | varible | — |
| IgD | — | — | |
| IgE | — | — | |
| IgG1 | ++++ | ++++ | |
| IgG2 | ++++ | ++++ | |
| IgG3 | — | ++++ | |
| IgG4 | ++++ | ++++ | |
| IgGM | varible | — | |
| Avian egg yolk | IgY | — | — |
| Cow | ++ | ++++ | |
| Dog | ++ | + | |
| Goat | — | ++ | |
| Guinea pig | IgG1 | ++++ | ++ |
| IgG2 | ++++ | ++ | |
| Hamster | + | ++ | |
| Horse | ++ | ++++ | |
| Koala | — | + | |
| Llama | — | + | |
| Monkey(rhesus) | ++++ | ++++ | |
| Mouse | IgG1 | + | ++++ |
| IgG2a | ++++ | ++++ | |
| IgG2b | +++ | +++ | |
| IgG3 | ++ | +++ | |
| IgM | variable | — | |
| Pig | +++ | +++ | |
| Rabbit | no distinction | ++++ | +++ |
| Rat | IgG1 | — | + |
| IgG2a | — | ++++ | |
| IgG2b | — | ++ | |
| IgG3 | + | ++ | |
| Sheep | +/- | ++ |
++++=strong binding; ++= medium binding; —=weak binding or no binding
| Problem | Possible Cause | Solution |
|---|---|---|
| The flow rate of the column is very low. | The sieve plate is blocked. | Clean or replace the sieve plate. |
| Column is clogged. | Cleaning in place(part 3). | |
| Filtering the sample solution by passing them through a 0.22μm or 0.45μm filter. | ||
| The curve is not stable during sample purification | Tiny air bubbles from buffer or sample. | De-gas buffers and samples. Do not allow the column to dry. |
| A considerable amount of sample has been loaded, but no specific antibody of interest is detected. | The concentration of antibody of interest is very low. | Purify the antibody using the specific antigen coupled to a beads (i.e., PabPur Sufolink Beads, Cat. No. SA018005). |
| The antibody is sensitive to low-pH elution buffer | Neutralize the eluted fractions with Neutralization Buffer immediately after elution. | |
| The IgG subclass does not bind to protein A. | Try other affinity chromatography media to purify the antibody, such as rProtein G Beads or rProtein A/G Beads. | |
| The recovery rate gradually decreases. | The sample is overloaded. | Reduce the loading volume. |
| The column is too dirty and the binding capicity is reduced. | Cleaning in place(part 3). |
· Ordering Information
| Product | Cat. No. | Size |
|---|---|---|
| rProtein A Beads | SA012005 | 5 ml |
| SA012025 | 25 ml | |
| SA012100 | 100 ml | |
| SA012500 | 500 ml | |
| SA01201L | 1 L | |
| SA01210L | 10 L | |
| AbPur rProtein A Kit | SA012K03 | 3T |
| rProtein A Beads 4FF | SA015005 | 5 ml |
| SA015025 | 25 ml | |
| SA015100 | 100 ml | |
| SA015500 | 500 ml | |
| SA01501L | 1 L | |
| SA01510L | 10 L | |
| AbCap A 4FF | SA015C11 | 1 X 1 ml |
| SA015C51 | 5 X 1 ml | |
| SA015C15 | 1 X 5 ml | |
| SA015C55 | 5 X 5 ml | |
| SA015CS | 3X1 ml+1X5 ml |
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