Rat IgG1 kappa Isotype Control [Alexa Fluor® 700]

Fragmentation in protein-based molecules continues to be a challenge during manufacturing and storage, and requires an appropriate control strategy to ensure purity and integrity of the drug product. Electrophoretic and chromatographic methods are commonly used for monitoring the fragments. However, size-exclusion chromatography often suffers from low resolution of low molecular weight fragments. Electrophoretic methods like CE-SDS are not compatible with enriching fragments for additional characterization tests such as MS. These limitations may result in inadequate control strategy for monitoring and characterizing fragments for protein-based molecules. Capillary western blotting was used in this study as an orthogonal method for characterization of fragments in an IgG1 antibody under reduced conditions. To achieve a comprehensive mapping of various fragments generated by thermal stress, capillary western profiles were generated using recognition antibodies for IgG kappa (kappa) light chain, Fc, and Fab regions that enabled unambiguous fragment identification. Additionally, three different enzymatic digestion methods (IdeS, PNGase F, and IgdE) were applied coupled with capillary western blotting for clip identifications. Finally, complementary data collected using traditional chromatographic and electrophoretic methods allowed to establish a comparison of analytical profiles with an added benefit of fragment identification offered by capillary western profiling. In addition to various Fc and Fab-related low molecular weight fragments, a non-reducible thio-ether linked 75 kDa HL fragment was also identified.

We investigated the antiasthmatic effect of mogroside V (Mog V) in mice with ovalbumin (OVA)-induced asthma. Administration of Mog V effectively attenuated OVA-induced airway hyperresponsiveness and reduced the number of inflammatory cells in bronchoalveolar lavage fluid (BALF). Histological examination showed that Mog V reduced the inflammatory infiltration of the lungs in the asthmatic mice. ELISAs suggested that Mog V effectively decreased the levels of IL-4, IL-5, and IL-13 in BALF and serum levels of OVA-specific IgE and IgG1 in the asthmatic mice. A quantitative reverse-transcription PCR assay also indicated that Mog V decreased the mRNA levels of IL-17A, IL-23, and ROR gamma t in the lungs of the asthmatic mice (the opposite effect on Foxp3 mRNA). Furthermore, Mog V significantly reduced the OVA-induced activation of NF-kappa B in the lungs. This study indicates that Mog V alleviates OVA-induced inflammation in airways, and this effect is associated with a reduction in NF-kappa B activation. Practical applications A traditional Chinese medicine herb has been reported to have a strong curative effect on asthma in clinical practice. Siraitia grosvenorii is known in China as a functional food product with the ability to improve lung function. Mogroside V is a triterpene glycoside isolated from S. grosvenorii. Nonetheless, the antiasthmatic effect of mogroside V has not been evaluated yet. The aim of this study was to investigate the antiasthmatic activity of mogroside V in mice with chemically induced asthma. The data from this study will provide some scientific evidence supporting wider use of S. grosvenorii in functional foods.