Adiponectin


Description

Adiponectin is a hormone produced by the adipose tissue involved in the regulation of glucose and fatty acid oxidation. Serum adiponectin correlates with decreased free fatty acid blood concentrations and reduced body mass index or body weight; adiponectin protects from vascular diseases by inhibiting local proinflammatory signals, preventing preatherogenic plaque formation, and by impeding arterial wall thickening (Schondorf, Maiworm et al. 2005). Modulation of the circulating adiponectin concentration is observed in obesity-related disorders such as insulin resistance, type 2 diabetes mellitus and atherosclerosis, and it also occur in pathologies such as cancer and rheumatoid arthritis (Brochu-Gaudreau, Rehfeldt et al. 2010). We have not been able to find reagents that recognize MAR adiponectin.


Alignment

Protein alignment for human, rhesus macaque and marmoset adiponectin:

The referenced media source is missing and needs to be re-embedded.

 

Protein alignment for marmoset, owl monkey, and squirrel monkey adiponectin:

The referenced media source is missing and needs to be re-embedded.

 

References

  • Brochu-Gaudreau, K., C. Rehfeldt, R. Blouin, V. Bordignon, B. D. Murphy and M. F. Palin (2010). "Adiponectin action from head to toe." Endocrine 37(1): 11-32.
  • Schondorf, T., A. Maiworm, N. Emmison, T. Forst and A. Pfutzner (2005). "Biological background and role of adiponectin as marker for insulin resistance and cardiovascular risk." Clin Lab 51(9-10): 489-494.

 

Status

Marmoset adiponectin was produced in HEK293 cells with immunogenicity tags. A total of six clones were selected after screening using ELISA, FluoroSpot (for clonality analysis) and Octet (for affinity scouting and epitope binning). Clones were amplified on a small scale and mAbs were produced as pure or biotinylated molecules. Different capture/detection mAb pairs were tested for reactivity with recombinant MAR Adiponectin, or reactivity with marmoset plasma (Figure 1). 

NWMIR-ELISA screening of capture/detection mAb pairs specific for MAR Adiponectin.
Figure 1. ELISA screening of capture/detection mAb pairs specific for MAR Adiponectin.

The ELISA results suggested that DP2 and DP15 were the only capture antibodies able to bind Adiponectin from marmoset samples. Since Adiponectin is a multimeric protein, the same antibody was able to function as capture and detection (that is DP2 capture/DP2 detection, or DP15 capture/DP2 detection), but the DP2 capture/DP15 detection showed higher signals.

Another ELISA was set up to determine whether mAb pairs targeting different epitopes (DP2 capture/DP40 detection) will be better than the DP2 capture/DP15 detection pair, which target the same Adiponectin epitope (Figure 2). Results from this assay showed that, even though DP15 and DP40 target the same epitope, they produce the highest signals for Adiponectin molecules from the three NWM species tested. 

NWMIR-ELISA assay for MAR Adiponectin
Figure 2. ELISA assay for MAR Adiponectin. The DP2 mAb was used as capture Ab and tested against DP15 or DP40 detection antibodies. Reactivity was tested with plasma samples from all three NWM species.

In summary, we have generated mAbs that bind native MAR Adiponectin and identified mAbs pairs that work in ELISA format for marmoset, squirrel monkey, and owl monkey. The DP2/DP15 mAb pair will be tested on the LMX platform.