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New standard resins for antibody purification
KANEKA KanCapA™

Highly cross-linked cellulose bead

KANEKA KanCapA™ is an affinity chromatography resin immobilized Protein A.  It was designed for the purification of biomolecules containing an Fc region of immunoglobulin and its derivatives derived from biological samples and cell culture media.  KANEKA KanCapA™ is composed of highly cross-linked cellulose beads and Alkaline Resistant Protein A ligand.  It can be used for affinity purification of IgG at development, pilot, etc.


1. Link

2. Features

  • Alkaline resistant
  • Optimization of the eluting condition
  • High antibody binding capacity
  • Recyclable
  • Scale up is easy.

3. Recombinant protein A

Ligand of KanCapA™ is adopted for new type of Protein A containing 5-mer of mutant C domain not wild type Protein A. It has been reported that C domain of wild type Protein A bind to Fc domain as well as Fab domain. Especially, it has high affinity to an antibody of VH3 subfamily. In order to dissociate this type of binding, elution at low acidic conditions is required. In order to improve these problems, a new type Protein A was developed. This protein was newly designed by molecular design system and alkali-resistant. The problem of nonspecific binding to Fab domain and elution profile has been improved.

                        Protein A adopted in the product:

4. Product Characteristics

    Matrix     Highly cross-linked cellulose
    Mean particle size 1)     65 ~ 85 mm
    Ligand     Protein A, recombinant (Alkali resistant)
    Bond     Chemical bond (reductive amination)
    Dynamic Binding Capacity 2)     ≥ 35 mg human polyclonal IgG/mL carrier
    Chemical stability 3)     Stable under universal solvent for affinity chromatography  
    Range of pH 3)     pH 2 ~ 13
    Condition for cleaing in place 3)     0.1 ~ 0.5 mol/L Sodium hydroxide
    6 ~ 8 mol/L Urea, 6 mol/L Guanidine hydrochloride
    Flow rate     Maximum 500 cm/h (Bed height: 20 ~ 25 cm)
    Resident time     ≥ 3 minutes (Recommendation: 4 ~ 6 minutes)
    Storage conditions 4)     20% Ethanol suspension, 1 ~ 10°C


1)   Mean particle size is a middle one in the accumulated volume distribution.
2)   5 % Dynamic Binding Capacity is measured by frontal analysis (Residence time 3 minutes).
3)   It can be washed under alkaline conditions, but please avoid long-term washing operation.
4)   Do not freeze.

5. Fab region purification application

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        Utilizing the feature of KANEKA KanCapA™, Fc region and Fab region can be fractionated from IgG solution after papain treatment.


6. IgG eluion profile (pH 3.5 and pH 3.0)

image

        Since elution range of KANEKA KanCapA™ has been optimized, sharp fractionation of purified fraction is possible.

7. Recycle of resin by alkali washing

image

Profile data of IgG avidity after recycle treatment
with 0.1M sodium hydroxide solution

image

Profile data of IgG recovery rate and leaching
volume of protein A ligand by recycle treatment
(treated with 0.1M sodium hydroxide solution for 15 min.)

        High recycle efficiency has been confirmed as the reduction rate of IgG binding capacity is under 20% even after 300 times
              of the recycle treatment (treated with 0.1M sodium hydroxide solution for 15 minutes).

8. Dynamic binding capacity profile data

image

Dynamic Binding Capacity (DBC)* of KANEKA KanCapA™ Profile data

        KANEKA KanCapA™ has been confirmed to show high DBC in 4 ~ 6 minutes of the residence time.

* Dynamic Binding Capacity is defined as a value that indicates recovery efficiency under the samples to be purified are flowing.
  Since a carrier with high DBC can recover a lot of target molecules under high flow rate, it can be said as a superior carrier that can purify the target
  molecules efficiently in a short time of the recycle treatment (treated with 0.1M sodium hydroxide solution for 15 minutes).

9. Plot data of pressure / flow by sizes of resin filled column

image

    The filling floor-dependent pressure is calculated by deducting the system pressure from the total pressure.

10. Filling efficiency by column of various sizes

Filling method L.D. (cm) Height (mm) Plate / m Asymmetry
Flow filling
4.4 200 5710 1.14
Axial force filling
7 199 6063 1.06
Flow filling
10 201 5276 1.18
Flow filling
30 200 5208 1.17
Axial force filling
45 233 4712 1.09
Axial force filling
60 250 6257 1.06

    The data is made by the pulse injection technique under the following conditions.
    Solvent: 1 (v/v) % Acetone
    Injection volume: 1% column capacity
    Flow rate: 60 cm/h

        KANEKA KanCapA™ has been confirmed to be used by being filled in columns of various sizes.

11. Product List

Product Name Package Size Wako Catalog No. Grade Storage Condition
    KANEKA KanCapA™
  2 mL   (net  1 mL)
10 mL   (net  5 mL)
50 mL   (net 25 mL)
114-01071
110-01073
118-01074
for Genetic Research Keep at 2-10°C.
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