Quantitative determination of humanized monoclonal antibody rhuMAb2H7 in cynomolgus monkey serum using a Generic Immunoglobulin Pharmacokinetic (GRIP) assay.

Preclinical pharmacokinetic (PK) assays are important to help evaluate the safety and efficacy of a potential biotherapeutic before clinical studies. The assay typically requires a biotherapeutic-specific reagent to minimize matrix effects especially when the host species are non-human primates such as cynomolgus monkeys and the biotherapeutic is a humanized monoclonal antibody (MAb). Recombinant humanized mAb 2H7 (rhuMAb2H7) binds to the extracellular domain of CD20 that is expressed on B cells and results in B cell depletion. It is currently being evaluated for its therapeutic potential in rheumatoid arthritis (RA) in clinical studies. During the early development of rhuMAb2H7, a cynomolgus monkey PK assay was needed to help assess the pharmacokinetic parameters of rhuMAb2H7 in a pilot cynomolgus monkey study. However, development of a cynomolgus monkey PK assay was challenging due to lack of rhuMAb2H7-specific reagents. Here we describe an alternative method for detection of rhuMAb2H7 in cynomolgus monkey serum using polyclonal antibodies against human IgGs. This assay quantifies rhuMAb2H7 in 10% cynomolgus monkey serum with high sensitivity, accuracy, and precision. This assay successfully supported the rhuMAb2H7 development, and has the potential to be used to quantify other humanized MAb biotherapeutics in serum from a variety of non-human species.

Bioanalytical challenges and improved detection of circulating levels of IL-13.

BACKGROUND: IL-13 is a key mediator of type 2 inflammation-driven diseases. Circulating IL-13 levels are very low and challenging to detect reliably. We assessed the ability of immunoassays on the Erenna(®) and IMPACT platforms to measure serum IL-13 in asthma, idiopathic pulmonary fibrosis (IPF) and atopic dermatitis (AD) patients and in healthy controls (HC). RESULTS: The Erenna IL-13 assay exhibited significant specificity issues and had limited ability to detect IL-13 in serum samples. The IMPACT IL-13 assay had excellent specificity and detected IL-13 in 100% of serum samples tested from asthma, IPF and AD patients and HC. Serum IL-13 levels were significantly elevated in asthma, IPF and AD patients, relative to HC. CONCLUSION: The IMPACT IL-13 assay had fg/ml sensitivity and excellent specificity, enabling reliable detection of circulating levels of IL-13.

Characterization of IL-17AA and IL-17FF in rheumatoid arthritis and multiple sclerosis.

AIM: IL-17 is thought to play a prominent role in immune disorders. Sensitive and specific IL-17AA and IL-17FF assays were developed and used to determine levels in serum and cerebrospinal fluid (CSF) from patients with rheumatoid arthritis and relapsing remitting multiple sclerosis (RRMS). RESULTS: Qualified assays detected IL-17AA and IL-17FF in healthy and disease samples. Serum IL-17AA was significantly higher in rheumatoid arthritis and RRMS as compared with normal healthy subjects. IL-17AA was also elevated in RRMS CSF as compared with normal healthy subjects; although correlation was observed between serum levels of the two isoforms, no correlation was detected between serum and CSF levels. CONCLUSION: Reliable determination of IL-17 isoforms in the systemic and CNS compartments sheds light on the involvement of IL-17AA and IL-17FF in autoimmunity.