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| Product Name | Protein A Magnetic Beads |
| Catalog No. | MAGBEA-0010 |
| Description | Protein A-functionalized superparamagnetic beads designed for the rapid, high-capacity purification of IgG antibodies from serum, plasma, ascites fluid, cell culture supernatant, and other complex biological matrices. Recombinant Protein A, covalently conjugated through its C-terminus for optimal orientation of the IgG-binding domains, provides high-affinity capture of IgG subclasses from human, rabbit, mouse (IgG2a, IgG2b), guinea pig, and other species. The magnetic format enables gentle, non-denaturing antibody purification without column packing or centrifugation. |
| Intended Use | Small-scale to medium-scale purification of monoclonal and polyclonal IgG antibodies from cell culture harvest, ascites, serum, and plasma for biochemical characterization, labeling, fragmentation, and functional assay development. |
| Principle / Technology | A recombinant engineered variant of Staphylococcal Protein A (approximately 42 kDa) containing five IgG-binding domains is covalently coupled to the bead surface through lysine residues near the C-terminus using oriented thioether or amide linkage chemistry. This orientation exposes the N-terminal IgG-binding domains (E, D, A, B, C domains) for optimal antibody capture. IgG binds to Protein A at near-neutral pH through Fc region recognition, while contaminating host cell proteins, DNA, and media components are removed by washing. Bound IgG is eluted under mildly acidic conditions (pH 2.5-3.5) and immediately neutralized. |
| Detection Method | DLS for bead size; BCA assay for Protein A density; IgG binding isotherm analysis by UV280; SDS-PAGE and SEC-HPLC for purified antibody characterization; ELISA for IgG subclass specificity; endotoxin testing; Protein A leaching ELISA |
| Sample Type | Monoclonal and polyclonal IgG from: human (IgG1, IgG2, IgG4), rabbit (all subclasses), mouse (IgG2a, IgG2b, IgG3), guinea pig, goat, bovine, horse; human IgG3 binds weakly; mouse IgG1, rat IgG, and chicken IgY have negligible binding by Protein A; cell culture supernatants (CHO, HEK293, hybridoma), serum, plasma, ascites fluid |
| Performance Range / Specifications | Particle diameter: 1-2 µm; Protein A density: 5-15 mg Protein A per mL settled beads; IgG binding capacity: 20-40 mg human IgG per mL settled beads; dynamic binding capacity: 15-30 mg/mL at 10-minute residence time equivalent; non-specific binding: <5% of total bound protein; IgG purity: >95% by SDS-PAGE; Protein A leaching: <10 ng/mg IgG eluted; magnetic separation: <10 seconds; recommended binding pH: 7.0-8.0; elution pH: 2.5-3.5 |
| Sensitivity / LOD | Detectable IgG purification from solutions containing <10 µg/mL antibody; small-scale purification (microgram scale) reproducible with micro-batch protocol; IgG quantification linear from 0.1-10 mg/mL loading range |
| Specificity | Protein A binds IgG through specific Fc region interaction (CH2-CH3 domain interface); no binding of IgM, IgA, IgE, or IgD; minimal binding of Fab and F(ab')2 fragments; host cell protein reduction >100-fold in single pass from cell culture supernatant; no binding of albumin, transferrin, or other serum proteins under recommended conditions |
| Reaction Conditions / Protocol | Equilibrate beads with binding buffer (PBS pH 7.4 or Tris pH 8.0); add antibody-containing sample; incubate at room temperature for 10-30 minutes with end-over-end rotation; magnetically separate beads; wash 3 times with binding buffer; elute with 0.1 M glycine-HCl pH 2.5-3.0 or 0.1 M citrate pH 3.0; incubate 2-5 minutes; separate beads magnetically; immediately neutralize eluate with 1/10 volume 1 M Tris-HCl pH 8.5-9.0; total protocol: 30-60 minutes; beads can be regenerated 5-10 times with proper cleaning |
| Components / Formulation | Protein A-coated superparamagnetic beads, 25% or 50% slurry in PBS pH 7.4 with 20% ethanol or 0.02% sodium azide as preservative; recombinant Protein A (E. coli expressed, engineering for enhanced stability and reduced non-specific binding); binding/wash and elution buffers not included |
| Storage Conditions | 2-8°C in 20% ethanol (long-term) or PBS with sodium azide; do not freeze (freezing denatures Protein A); protect from light; store upright; for short-term use (<1 month), PBS with azide at 2-8°C is acceptable; always store in preservative-containing buffer |
| Shelf Life | 24 months at 2-8°C in 20% ethanol; 12 months in PBS with sodium azide at 2-8°C; Protein A activity >90% retained through shelf life when stored as recommended; reduced shelf life after first use due to microbial contamination risk |
| Package Specifications | 1 mL, 5 mL, 10 mL, 25 mL settled bead volume (25-50% slurry); supplied as slurry in storage buffer; convenient pre-packed columns also available; bulk OEM quantities available |
| Product Form | Dark brown to black suspension in storage buffer; homogeneous after gentle shaking (do not vortex vigorously with ethanol—flammable); beads settle rapidly from slurry; supernatant clear after magnetic separation; consistent appearance across lots |
| Quality Control | Per lot: Protein A density (BCA and UV), IgG binding capacity (human polyclonal IgG isotherm), IgG subclass specificity panel (ELISA), purity of eluted IgG (>95% by SDS-PAGE), Protein A leaching (<10 ng/mg IgG by ELISA), endotoxin (<0.1 EU/mg), sterility, magnetic separation time; functional test with CHO cell culture supernatant |
| Key Features | Oriented Protein A coupling for maximum IgG-binding domain accessibility; high binding capacity (20-40 mg/mL); low Protein A leaching; recombinant Protein A with engineered alkaline stability (tolerates 50+ CIP cycles with 0.1 M NaOH); magnetic format eliminates columns and pumps; rapid batch binding protocol; compatible with multiple species IgGs |
| Purity | Protein A >95% by SDS-PAGE; no detectable protease activity; IgG eluted from beads >95% purity by SDS-PAGE and SEC-HPLC; endotoxin <0.1 EU/mg; host cell protein in eluted IgG <500 ppm; residual DNA <10 pg/mg IgG |
| Concentration | 25% or 50% slurry (v/v settled beads in storage buffer); Protein A 5-15 mg/mL settled beads; exact concentration and IgG capacity in lot certificate; bead count approximately 10^9-10^10 particles per mL settled beads |
| Activity / Unit Definition | Static IgG binding capacity: 20-40 mg human IgG/mL settled beads; dynamic binding capacity: 15-30 mg/mL (10-min residence equivalent); binding constant (Kd): ~10^-7 to 10^-8 M for human IgG1; binding capacity retained >90% over 5-10 regeneration cycles |
| Molecular Weight | Recombinant Protein A: ~42 kDa (engineered variant with 5 IgG-binding domains); bead molecular weight not applicable |
| Source / Origin | Recombinant Protein A expressed in E. coli with engineered C-terminal cysteine for oriented coupling; magnetic beads are fully synthetic polymer-magnetite composite; all components synthetic or recombinant; certified animal-origin-free; BSE/TSE free; no Staphylococcal culture used |
| pH Range / Optimal pH | IgG binding: pH 6.5-9.0 (optimal pH 7.0-8.0); IgG elution: pH 2.5-3.5; bead stability: pH 2-12; NaOH cleaning (0.1 M NaOH, 15 minutes): tolerated; long-term storage: 20% ethanol preferred or PBS with sodium azide at pH 7.4 |
| Shipping Conditions | Ambient temperature in 20% ethanol; cold packs optional; do not freeze; 20% ethanol is flammable liquid—appropriate shipping classification required for air transport; water-based (PBS/azide) formulation: ambient or cold packs, no dangerous goods |
| Expiration Date / Stability | 24 months in 20% ethanol at 2-8°C; 12 months in PBS/azide at 2-8°C; Protein A activity verified at 0, 12, 24 months; no increase in Protein A leaching over time; ethanol concentration maintained during storage; opened container stability dependent on handling practices |
| Regulatory / Compliance | For research use and further manufacturing; suitable for purification of antibodies for preclinical research; not for production of therapeutic antibodies without additional regulatory filings; recombinant product with full traceability; ISO 9001 manufacturing; DMF support available |
| Compatibility | Compatible with all common binding buffers: PBS, Tris, HEPES (20-100 mM, pH 7.0-8.0); elution buffers: 0.1 M glycine-HCl pH 2.5-3.0, 0.1 M citrate pH 3.0-3.5, 0.1 M acetic acid pH 2.8; neutralization: 1 M Tris-HCl pH 8.5-9.0 (add 1/10 volume); cleaning: 0.1 M NaOH (15-minute contact), 6 M guanidine HCl (strips residual protein); avoid phosphate during NaOH cleaning (forms precipitate) |
| Recommended Buffer System | Storage: 20% ethanol in water or PBS pH 7.4 + 0.02% NaN3; binding: PBS pH 7.4 or 20 mM Tris pH 8.0, 150 mM NaCl; elution: 0.1 M glycine-HCl pH 2.5-3.0; neutralization buffer: 1 M Tris-HCl pH 9.0; cleaning: 0.1 M NaOH |
| Application Notes / Precautions | For serum/plasma samples, pre-filter or centrifuge to remove particulates before bead incubation; dilute viscous samples 1:1 with binding buffer for better bead contact; elution efficiency is improved by pre-warming elution buffer slightly (room temperature); Protein A binds human IgG1 and IgG2 with high affinity, IgG4 moderate; for mouse IgG1 purification, consider Protein G beads instead; after NaOH cleaning, re-equilibrate beads thoroughly before reuse; beads can be regenerated 5-10 times with proper cleaning and storage |
| Batch-to-Batch Consistency | Protein A density CV <12% between lots; IgG binding capacity CV <10%; Protein A leaching <10 ng/mg across all lots; eluted IgG purity >95% for all lots; endotoxin <0.1 EU/mg per lot; comprehensive lot characterization documented |
For research use only, not for clinical use.
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