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MonoRab™ Anti-Humanized VHH Affinity Resin FF

Figure 3. Reuse times of anti-humanized VHH affinity resin FF

After VHH was captured by MonoRab™ Rabbit Anti-Humanized VHH Affinity Resin FF, the resin was washed twice with TBS (10 times bed volume), and eluted with 10 times bed volume of acid elution buffer (pH 2.5). The resin was washed with TBS and elution buffer again. Then the resin was regenerated by 10 times bed volume of regeneration buffer A and regeneration buffer B. Binding of VHH was repeated 9 times, and the binding capacity did not decrease significantly, indicating that the resin can be reused at least 10 times.

MonoRab™ Anti-Humanized VHH Affinity Resin FF

Figure 1. Purification of Humanized VHH#1 protein from E. coli lysate

Lanes:
1. Protein marker
2. Whole cell lysate
3. Flow through
4. Eluate of target Humanized VHH single domain antibody-1 (M.W.=15 kDa)

MonoRab™ Anti-Humanized VHH Affinity Resin FF

Figure 2. Purification of Humanized VHH#2 protein from E. coli lysate

Lanes:
1. Protein marker
2. Whole cell lysate
3. Flow through
4. Eluate of target Humanized VHH single domain antibody-2 (M.W.=15 kDa)

MonoRab™ Anti-Humanized VHH Affinity Resin FF

The product is used for purification of humanized VHH single domain antibody from commonly used protein expression systems
L00951
¥3800

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Description

Camelid VHH Antibody (also called single-domain antibody, sdAb or Nanobody by Ablynx) is a peptide chain about 110 amino acids in length, containing one variable domain of a heavy-chain antibody (a VHH fragment). Like a whole antibody, the camelid VHH antibody is able to bind selectively to a specific antigen. This single-domain antibody has a broad range of applications in biotechnical and therapeutic research fields due to its small size, simple production and high affinity. However, their sequences may increase the risk of immunogenicity and anti-drug antibody (ADA) development in humans, and thus, sdAbs are routinely humanized during development.  
GenScript’s MonoRab™ Anti- Humanized VHH Affinity Resin FF (Cat. No. L00951)
is designed for purification of humanized VHH single domain antibody from commonly used protein expression systems such as bacteria, yeast and mammalian cells. The VHH single domain antibody in cell lysate can specifically bind to the Anti- Humanized VHH monoclonal antibody coupled to the resin. Nonspecific binding reactions are eliminated by stringent washing steps, and the purified target protein is eluted with a high recovery rate. Table 1 lists the main characteristics of GenScript’s MonoRab™ Anti- Humanized VHH Affinity Resin FF. 


Table 1. Characteristics of Anti-Humanized VHH Affinity Resin FF
Product content50% settled resin in TBS with 0.02% sodium azide
Matrix4% highly cross-linked agarose
Average bead size90 μm
LigandMonoRab™ Anti-Humanized VHH Antibody
Binding capacity> 2 mg Humanized VHH per mL settled resin (depending on amino acid sequence of different Humanized VHHs)
Storage and stabilityStore at 2-8 °C for up to 12 months. Do not freeze the resin.
Resin reuseWhen stored at 2 - 8° C, the resin can be recycled at least 5 times with no loss of binding capability. While, it can be reused up to 10 times with minimum loss of binding capability.
Elution methodAcid buffer elution (pH 2.5)
Reagents compatibilityCompatible with commonly used bacterial lysis reagents. For reagents compatible with purifications, see tables 2 and 3 in the manual. 

Note

Limited Use Label license: This product may be the subject of one or more patents filed by GenScript Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer should not sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. For commercial use, please contact GenScript at product@genscript.com.

For research use only. Not intended for human and animal therapeutic or diagnostic use.

  • MonoRab™ Anti-Humanized VHH Affinity Resin FF
  • MonoRab™ Anti-Humanized VHH Affinity Resin FF

    Figure 3. Reuse times of anti-humanized VHH affinity resin FF

    After VHH was captured by MonoRab™ Rabbit Anti-Humanized VHH Affinity Resin FF, the resin was washed twice with TBS (10 times bed volume), and eluted with 10 times bed volume of acid elution buffer (pH 2.5). The resin was washed with TBS and elution buffer again. Then the resin was regenerated by 10 times bed volume of regeneration buffer A and regeneration buffer B. Binding of VHH was repeated 9 times, and the binding capacity did not decrease significantly, indicating that the resin can be reused at least 10 times.

  • MonoRab™ Anti-Humanized VHH Affinity Resin FF
  • MonoRab™ Anti-Humanized VHH Affinity Resin FF

    Figure 1. Purification of Humanized VHH#1 protein from E. coli lysate

    Lanes:
    1. Protein marker
    2. Whole cell lysate
    3. Flow through
    4. Eluate of target Humanized VHH single domain antibody-1 (M.W.=15 kDa)

  • MonoRab™ Anti-Humanized VHH Affinity Resin FF
  • MonoRab™ Anti-Humanized VHH Affinity Resin FF

    Figure 2. Purification of Humanized VHH#2 protein from E. coli lysate

    Lanes:
    1. Protein marker
    2. Whole cell lysate
    3. Flow through
    4. Eluate of target Humanized VHH single domain antibody-2 (M.W.=15 kDa)


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