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purification resins

Protein A, G and L Resins

GenScript offers recombinant protein A with non-essential domains removed, recombinant protein G with albumin binding site deleted as well as codon optimized protein L. By pre-coupling these genetically modified proteins to 4% highly cross-linked agarose, GenScrtipt developed excellent antibody affinity chromatography matrix - Protein A, G and L Resins for batch/gravity purification of polyclonal and monoclonal antibody from cell culture supernatants, serum, and ascites at both laboratory and larger scale. With enhanced binding capacity, these resins enable you to get highly purified antibodies with minimal contaminations, resulting in cleaner preparations and greater yields.

Highlights



Protein A, protein G and protein L are surface proteins originally found in cell wall of bacteria Staphylococcus aureus, Group G Streptococci and Peptostreptococcus magnus, respectively. Protein A and protein G bind with high affinity and specificity to Fc region of immunoglobulins from many of mammalian species, most notably IgGs. Unlike protein A and G, Protein L binds immunoglobulins through kappa light chain. These proteins have been usually used to purify, immobilize or detect immunoglobulins.

Ordering Information

Cat. No.
Product Name
Size
Price
Quantity
L00433-55 Monofinity A Resin Prepacked Column 5mlx5
5 ml prepacked x 5
$1809.00
L00433-51 Monofinity A Resin Prepacked Column 5mlx1
5 ml prepacked x 1
$499.00
L00433-5 Monofinity A Resin
5 L
L00433-25 Monofinity A Resin
25 ml
$1680.00
L00433-200 Monofinity A Resin
200 ml
$3350.00
L00433-15 Monofinity A Resin Prepacked Column 1mlx5
1 ml prepacked x 5
$549.00
L00433-11 Monofinity A Resin Prepacked Column 1mlx1
1 ml prepacked x 1
$169.00
L00433-10 Monofinity A Resin
10 L
L00433-1 Monofinity A Resin
1 L
L00210 Protein A Resin
5 ml
$98.00
L00464-5 Protein A Resin FF
5 ml
$390.00
L00209 Protein G Resin
5 ml
$129.00
L00239 Protein L Resin
2.5 ml
$145.00
L00664-5 Protein G Resin FF
5 ml
$416.00
L00664-25 Protein G Resin FF
25 ml
$1473.00
L00664-100 Protein G Resin FF
100 ml
$2209.00
L00680-55 Protein A Resin FF Prepacked Column
5 ml x 5
$2083.00
L00680-51 Protein A Resin FF Prepacked Column
5 ml x 1
$465.00
L00680-12 Protein A Resin FF Prepacked Column
1 ml x 2
$251.00
L00681-55 Protein G Resin FF Prepacked Column
5 ml x 5
$3410.00
L00681-51 Protein G Resin FF Prepacked Column
5 ml x 1
$761.00
L00681-12 Protein G Resin FF Prepacked Column
1 ml x 2
$411.00
 


Table 1. Properties of the recombinant Protein A, G and L on Resins

Protein A Protein G Protein L
Source Staphylococcus aureus Streptococcus Peptostreptoccus magnus
Molecular weight 34 kDa 22 kDa 42kDa
Numbers of IgG(Ig) binding sites 5 3 5
IgG Binding capacity >20 mg human IgG/ml settled resin >20 mg sheep IgG per ml settled resin >15 mg rabbit Ig per ml settled resin

Table 2. Selection Guide of Protein A, G and L Resins

 
Monofinity A
Protein A Resin
Ultra Protein A Resin
Protein A Resin FF
Protein G Resin
Protein L Resin
Matrix
Highly crosslinked 4% beaded agarose
Agarose, 4%
Agarose, 4% crosslinked
Highly crosslinked 4% beaded agarose
Agarose, 4% crosslinked
Agarose, 4% crosslinked
Average bead size
~ 90um
~ 90um
~ 90um
~ 90um
~ 90um
~ 90um
Ligand
Alkali-Tolerant Protein A
Recombinant Streptococcal protein A expressed in
E. coli
Recombinant Streptococcal protein A expressed in
E. coli
Recombinant Streptococcal protein A expressed in
E. coli
Recombinant Streptococcal Protein G lacking the albumin-binding produced in
E. coli
Highly purified protein L
Binding Capacity
> 30 mg hIgG/ml resin
> 20 mg human IgG/ml resin
30 - 40 mg hIgG /ml settled resin
> 40 mg rabbit IgG/ml settled resin
>20 mg sheep IgG/ml settled resin
>15 mg rabbit Ig/ml settled resin
Chemical Stability
0.5M NaOH for CIP
Stable with all commonly used reagents
Stable with all commonly used reagents
Stable with all commonly used reagents
Stable with all commonly used reagents
Stable with all commonly used reagents
Stable with all commonly used reagents
Working pH
3-12
3-9
3-9
3-9
3-9
2 to 10
Clean in place
0.1-0.5M NaOH
6 M guanidine hydrochloride
0.1% Triton™ X-100
Alternatively, wash the column with 70% ethanol
6 M guanidine hydrochloride
0.1% Triton™ X-100
Alternatively, wash the column with 70% ethanol
6 M guanidine hydrochloride
0.1% Triton™ X-100
Alternatively, wash the column with 70% ethanol
0.1% Triton™ X-100
Alternatively, wash the column with 70% ethanol
Washing of the packed column with 70% ethanol
15mM NaOH
Linear flow velocity
50-300cm/h
50-300cm/h
50-300cm/h
50-300cm/h
50-300cm/h
50-300cm/h
Storage
20% ethanol, 2 - 8°C
20% ethanol, 2 - 8°C
1X PBS containing 20% ethanol, 2 - 8°C
1X PBS containing 20% ethanol, 2 - 8°C
1X PBS containing 20% ethanol, 2 - 8°C
1X PBS containing 20% ethanol, 2 - 8°C

Figure 1. GenScript protein A Resin has about 16% higher DBC10% than the competitor's at Retention Time of 6 minutes, and about 10% higher SBC than the competitor's



Figure 2. Affinity purification of rabbit IgG from rabbit serum, using Protein A Resin (L00210)


Lane 1: Protein standard;
Lane 2: Rabbit serum;
Lane 3: Flow-through of serum;
Lane 4: Wash;
Lane 5: Eluted rabbit IgG (50 kDa and 25 kDa).

Citations

  • Tradtrantip L., et al. Therapeutic Cleavage of Anti–Aquaporin-4 Autoantibody in Neuromyelitis Optica by an IgG-Selective Proteinase. Mol Pharmacol. 2013 Jun;83(6):1268-75.
  • Song SE., et al. IGFBP5 mediates high glucose-induced cardiac fibroblast activation. J Mol Endocrinol. 2013 Apr 12;50(3):291-303.
  • Dong DJ., et al. Steroid hormone 20-hydroxyecdysone regulation of the very-high-density lipoprotein (VHDL) receptor phosphorylation for VHDL uptake. Insect Biochem Mol Biol. 2013 Apr;43(4):328-35.
  • Yin Y., et al. Differentially Expressed Genes of Human Microvascular Endothelial Cells in Response to Anti-Dengue Virus NS1 Antibodies by Suppression Subtractive Hybridization. Viral Immunol. 2013 Jun;26(3):185-91.
  • Jennings VA., et al. Lymphokine-activated killer and dendritic cell carriage enhances oncolytic reovirus therapy for ovarian cancer by overcoming antibody neutralization in ascites. Int J Cancer. 2014 Mar1;134(5):1091-101.
  • Hwang JH., et al. Protein Phosphatase 2A Isoforms Utilizing Aβ Scaffolds Regulate Differentiation through Control of Akt. J Biol Chem. 2013 Nov 1;288(44):32064-73.
  • Lan JF., et al.Prohibitin interacts with envelope proteins of white spot syndrome virus and prevents infection in red swamp crayfish Procambarus clarkii. J Virol. 2013 Dec;87(23):12756-65.
  • Asavapanumas N. et al. Unique neuromyelitis optica pathology produced in naïve rats by intracerebral administration of NMO-IgG. Acta Neuropathol. 2013 Nov 5.
  • Liu W., et al. The hormone-dependent function of Hsp90 in the crosstalk between 20-hydroxyecdysone and juvenile hormone signaling pathways in insects is determined by differential phosphorylation and protein interactions. Biochim Biophys Acta. 2013 Nov;1830(11):5184-92.
  • Mabashi-Asazuma H., et al. A novel baculovirus vector for the production of non-fucosylated recombinant glycoproteins in insect cells.Glycobiology. 2013 Dec 9.
  • XW Wang, et al. C-type lectin binds to β-integrin to promote hemocytic phagocytosis in an invertebrate. J Biol Chem. 2013 Dec 9.
  • Tubon TC Jr., et al. dCREB2-Mediated Enhancement of Memory Formation.J Neurosci. 2013 Apr 24;33(17):7475-87.
  • Short KR., et al. Antibodies mediate the formation of neutrophil extracellular traps (NETs) in the middle ear and facilitate secondary pneumococcal otitis media. Infect Immun. 2014 Jan;82(1):364-70.
  • Armistead JS., et al. Antibodies to a single, conserved epitope in Anopheles APN1 inhibit universal transmission of falciparum and vivax malaria. Infect Immun. 2013Dec 9.
  • Zhang D., et al. Generation and Characterization of a Novel Recombinant Antibody against LMP1-TES1 of Epstein-Barr Virus Isolated by Phage Display. Viruses. 2013 Apr 22;5(4):1131-42.
  • De Oliveira SN., et al. A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors. J Transl Med. 2013 Jan 29;11:23.
  • Safdari Y., et al. humMR1, a highly specific humanized single chain antibody for targeting EGFRvIII. Int Immunopharmacol. 2013 Dec 25;18(2):304-310.
  • Lin H., et al.A novel human Fab antibody for Trop2 inhibits breast cancer growth in vitro and in vivo.Int J Cancer. 2014 Mar 1;134(5):1239-49.
Tagged Protein Purification Resins: purification of polyhistidine, GST, biotin and streptavidin tagged proteins.
High-Affinity Iodoacetyl Resin and High-Affinity Antibody Purification Kit: covalent immobilization of ligands containing
  sulfhydryl groups for following affinity purification.
 
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