Separation Techniques

ANTIBODY PURIFICATION

 

  1. UptiSpin Protein A NEW

  2. UptiSpin Protein G NEW

  3. UptiSpin IMAC NEW

  4. Immobilized Protein A, HC

  5. Immobilized Protein A

  6. PBS

  7. Mild IgG elution buffer

  8. Immobilized Protein L

UptiSpin Protein A NEW           UptiSpin Protein G NEW           UptiSpin IMAC NEW

Affinity  Purification on protein A, G or on Nickel binded resins, of antibodies and recombinant proteins from serum, ascites and tissue culture supernatant derived from static cultures and bioreactors. 

Proteins are purified using a powerful affinity spin column ; a microfuge common to all biochemistry and immunology laboratories.

  • Convenient and easy to use

  • Fast

  • Cost effective : less than 1 Euro to purify 1 mg

  • Efficient : Proteins are sufficiently pure for further applications

Kits are provided with all necessary material (spin columns, centrifuge tubes, desalting columns, buffers...).

2 kits formats are available : Mini and Midi.

Binding capacity:

UptiSpin Protein A and Protein G:

Mini : 1 mg human IgG

Midi : 20 mg human IgG

UptiSpin IMAC

Mini : 1 mg His-tagged recombinant protein

Midi : 10 mg His-tagged recombinant protein

 

Cat.#

Description

 

 

 

 

 

PROTEIN A UptiSpin

MINI

UPBB9720

Protein A Mini Sample Kit : 2 A Plugs (buffers included)

UPBB9730

Protein A Mini Kit – 16 A Plugs (buffers included)

UPBB9740

Protein A Mini Bulk Pack – 48 A plugs (buffers not included)

MIDI

UPBB9750

Protein A Midi Kit – 4 Midi A Plugs (buffers included)

UPBB9760

Protein A Midi Bulk Pack – 12 A Plugs (buffers not  included)

TOP

PROTEIN G UptiSpin

MINI

UPBB9770

Protein G Mini Sample Kit : 2 G Plug (buffers included)

UPBA7770

Protein G Mini Kit – 16 G Plugs (buffers included)

UPBB9780

Protein G Mini Bulk Pack – 48 G plugs (buffer snot included)

MIDI

UPBB9790

Protein G Midi Kit – 4 Midi G Plugs (buffers included)

UPBB9800

Protein G Midi Bulk Pack – 12 G Plugs (buffers not included)

TOP

METAL CHELATE UptiSpin

MINI

UPBB9580

Metal Chelate Mini Sample Kit  - 8 Mini MC Plugs  (buffers included)

UPBB9600

Metal Chelate Mini Kit – 24 MC Plugs (buffers included)

UPBB9620

Metal Chelate Mini Pack – 24 MC plugs (buffers included, concentrators UF not included)

UPBB9630

Metal Chelate Mini Bulk Pack (buffers not included, concentrators UF not included)

MIDI

UPBB9640

Metal Chelate Midi Kit - 8 MC plugs (buffers included)

UPBB9650

Metal Chelate Midi Pack - 8 MC plugs (buffers included, concentrators UF not included)

UPBB9660

Metal Chelate Midi Bulk Pack – 24 MC Plugs (buffers not included, concentrators UF not included)

UPBB9640

Metal Chelate Midi Kit - 8 MC plugs (buffers included)

UPBB9650

Metal Chelate Midi Pack - 8 MC plugs (buffers included, concentrators UF not included)

TOP

 

Immobilized Protein A, HC     Immobilized Protein A     PBS     Mild IgG elution buffer     Immobilized Protein L

Antibody purification was popularized by Protein A affinity chromatography.

Uptima offers high-quality supports with Ig-binding proteins, Protein A and Protein L for the requirements from R&D level to production (ask for bulk sizes).

Choose the right support considering :

  • The specificity of the ligands,

  • Other parameters such as stability, leaching, flow rate…

Protein A suits particulary the purification of monoclonal antibodies from FCS supplemented cell cultures. It is often a first intention of choice for many other Ig species. Protein L is dedicated to specific applications especially Ig fragements.

Protein A and Protein L are available in classic grade, with unsurpassed binding capacity. Ligands are covalently bound to the matrice, a modified agarose, to ensure minimal leaching of ligand, excellent stability and to allow re-use.

-

Protein A

Protein L

(native)

(Staphylococcus)

(Peptostreptococcus)

Origin

E. coli

E. coli

Nb binding sites

(4) 4

4

Ig binding site

Fc

K light chain

Albumin binding

(-) -

+

Optimal pH binding

8-9

7.5

Binding capacity *

Hu IgGs

+++

+++

Hu IgG1

+++

+++

Hu IgG2

+++

+++

Hu IgG3

+

+++

Hu IgG4

+++

+++

Hu IgM

+

+++

Hu IgE

++

+++

Hu IgD

-

+++

Hu IgA

+

+++

Ms IgGs

+++

+++

Ms IgG1

+

+++

Ms IgG2a

+++

+++

Ms IgG2b

+++

+++

Ms IgG2c

+++

+++

Ms IgG3

+++

+++

Ms IgM

-

++

Rt IgGs

+

+++

Rt IgG1

+

++

Rt IgG2a

+++

+++

Rt IgG2b

+++

+++

Rt IgG3

+++

+++

Rb IgGs

+++

+

Gp IgGs

+++

+/-

Hs IgGs

++

+/-

Dog IgGs

+++

-

Cat IgGs

+++

-

Mk IgGs

+++

-

Pig IgGs

+++

+++

Gt IgGs

+/++

-

Sh IgGs

+/++

+

Bv IgGs

+/++

+/-

Dk IgGs

++

-

Hs IgGs

+

-

Ck IgGs

-

-

Bv : Bovine

Ck : Chicken

Ct : Cat

Dg : Dog

Gp : Guinea-pig

Gt : Goat

Hm : Hamster

Hu : Human

Hs : Horse

Mk : Monkey

Ms : Mouse

Rt : Rat

Rb : Rabbit

Sh : Sheep

-

* binding capacities for proteins L and LA are given for only K subclass

+++ : strong binding

-  : no binding

TOP

Immobilized Protein A, HC

UP904670

5 ml

Immobilized Protein A

UP49981G

2 ml

UP904672

20 ml

UP49981H 5 ml

The highest capacity support !

Protein A immobilized onto a 6% sepharose FF, supplied at 50% suspension

  • 30-40 mg of human IgG per ml of wet gel (static)

  • Excellent stability and minimal leaching of ligand

  • High flow rate, up to 400cm/hour

  • Pharmaceutical requirements

The classic and perfect choice for most applications

Protein A immobilized onto a 6% modified agarose, supplied at 50% suspension

  • Lowest leaching (<3ng Protein /mL)

  • 18-22 mg of human IgG per ml of wet gel

  • High flow rate, up to 300cm/hour

Applications

  • Purification of monoclonal antibodies, notably from FCS

  • Supplemented culture media (no Bovine IgGs binding)

  • Purification of polyclonal antibodies

  • Immunoprecipitation

  • IgG immobilization (affinity columns preparation)

  • Preadsorption, IgG depletion

  • Isolation of immune complexes

Technical Sheet

Applications

  • Purification of monoclonal antibodies, notably from FCS

  • Supplemented culture media (no Bovine IgGs binding)

  • Purification of polyclonal antibodies

  • Immunoprecipitation

  • IgG immobilization (affinity columns preparation)

  • Preadsorption, IgG depletion

  • Isolation of immune complexes

Technical Sheet

 

Cycle

Capacity (% initial)

Protein A in eluat

1

100%

2.5 ng/ml*

2

100%

2.4 ng/ml*

16

100%

1.5 ng/ml*

32

>95%

6.7 ng/ml*

100

>85%

3.3 ng/ml*

* Representative datas.

Eluat: polyclonal human IgG 7.4 - 8.5mg/ml

 

PBS

UP68723

Mild IgG elution buffer

UP38591

 

TOP

TOP

Protein A binds human IgG subclasses, IgM (medium), IgA and IgE, and mouse IgG1 (weakly), IgG2a and IgG2b. Protein A also binds IgGs from various other laboratory and domestic animals (+++/Rb, Mo, Pg, Dg, Ct, Ha, Gp, ++/Dk), but predominantly only isotypes from some animals (IgG2/Sh, Bv, Gt; IgG2c/Rt).

Uptima Protein A is produced by recombinant technology in E.coli, with culture media being exempt of any animal proteins. Highly controlled procedures give a material that is free from toxic bacterial contaminant (Staphyloccocus endotoxins and hemolysin), that are found normally in native Protein A.

The IgG1 purified fraction is :

  • >95% pure

  • Very concentrated

Immobilized Protein L

UP52746A

5 ml

Great for the purification of Ig fragments !

Protein L immobilized onto a 4% modified agarose, supplied at 50% suspension

  • 3-5 mg of human IgG per ml of wet gel

  • Ig binding does not interfere with antigen binding

  • Excellent stability

Applications

  • Ideal for the purification of Fab’2, Fab and scFv fragments containing kappa light chains

  • Great to purify kapa monoclonal antibodies directly from media supplemented with FCS

  • Purification of IgG, IgM, IgA, IgE, IgD containing kappa light chains from serum, ascites and hybridoma supernatant

  • Antibodies sensitive to alkaline pH

Technical Sheet

Immobilized Protein L, A

inquire

Joins the features of Protein A and protein L

Protein L (product #56874A) is a protein receptor from Peptostreptococcus magnus that binds to kappa light chains without interfering with antigen binding.

One protein L molecule contains 4 binding sites, like for protein A. Optimal binding occurs at pH 7.5.

Uptima recombinant protein L is expressed from E.coli.

For improved immunoprecipitations, or affinity purification, how to immobilize your antibody on Protein A et L ?

Use a cross-linker suitable for the reticulation of your Ab to the Ig binding protein.

DSS (UP28065) is a classic and efficient choice

 

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