Recommendations for the Bioanalytical Method Validation of Ligand-Binding Assays to Support Pharmacokinetic Assessments of Macromolecules

Abstract

In this consensus document, we attempt to make recommendations that are based on bioanalytical best practices and statistical thinking for development and validation of LBAs.

Pharmacokinetics and pharmacodynamics of ch14.18/CHO in relapsed/refractory high-risk neuroblastoma patients treated by long-term infusion in combination with IL-2

“…10 Experimental runs with CVs exceeding 20% were rejected from the analysis of PK parameters and subjected to reanalysis. Within-assay precision was determined by analysis of the CVs from eight samples containing known ch14.18/ CHO concentrations in human serum (0.6 and 0.3 mg/ml) ( Fig.…”

“…Instead of STI over five days (5 £ 20 mg/m2/d, 8h infusion), 53 high-risk NB patients received the same cumulative dose of 100 mg/m2 ch14.18/CHO given as a continuous long-term infusion (LTI) over 10 days (10£ 10 mg/m2/d). Similar to the reported randomized, open label Phase 3 clinical trial, 4 patients received ch14.18/CHO in combination with a cytokine (6£ 106/d IU/m2 subcutaneous (s.c.) interleukin (IL)-2, d 1-5, followed by a combined application of IL-2 (d [8][9][10][11][12] with LTI of ch14.18/CHO (d 8-18)). A reduced toxicity profile of the new treatment protocol was indicated by significantly decreased morphine usage and low pain scores.…”

“Fit-for-Purpose” Method Validation and Application of a Biomarker (C-terminal Telopeptides of Type 1 Collagen) in Denosumab Clinical Studies

“…It is recommended that a minimum of six non-zero concentrations be used when fitting a nonlinear (sigmoidal) concentration-response curve (8,13). We used the bulk stock reference material to prepare a ninepoint standard curve, including two anchor points, sufficient to define the four-parameter logistic function.…”

“…The accuracy and precision data were assessed using a validated accuracy and precision evaluation tool as described in the paper by DeSilva et al (8). The matrix comparison was performed during the accuracy and precision evaluation for this method.…”

“…The basic framework of method validation commonly used to support PK analyses in clinical studies was used as a basis for the method validation of CTx in human serum and to characterize assay accuracy, precision, sensitivity, selectivity, specificity, and stability (8). In addition, extensive validation experiments were conducted to support CTx as a biomarker in advanced application so that the data may be used for regulatory agency submission (9).…”