The sensitivity of bound and free κ and λ LC antisera in serum was approximately 100 mg/L (representative images not shown). All statistical analyses were conducted using PASW Statistics Version 18 (IBM, USA) with the exception of assay linearity, batch-to-batch variability and mAb correlations with Freelite™, which were assessed using the Microsoft Excel Add-in Analyse-it (version 2.26, www.analyse-it.com). Spearman correlations were ranked as ‘good’ between 0.75 and 0.90, and ‘excellent’ above 0.90. Olaparib manufacturer All figures were produced using
SigmaPlot version 11.0 (Systat Software Inc., USA). 250 plasma samples from healthy donors were analysed for κ and λ FLCs using the mAb assay and a comparison was made between each of the anti-FLC mAbs, and to Freelite™. All samples were pre-screened for paraproteins by routine serum IFE analysis. IFE revealed that one sample had an IgG λ paraprotein with λ FLC, and the sample was excluded from further analyses; both the mAb assay and Freelite™ assays identified elevated λ FLC and an abnormal κ:λ FLC
ratio in this sample. This finding accords with expected prevalence of MGUS in the general population (Kyle et al., 2006). Reference ranges for each anti-FLC mAb were similar to Freelite™ for the remaining 249 samples (Fig. 2). The two anti-κ mAbs also had similar reference ranges to each other, with BUCIS 04 having a slightly broader reference range than BUCIS 01 (BUCIS 01: 6.46–15.10 mg/L; BUCIS 04: 4.35–19.44 mg/L). The two anti-λ mAbs http://www.selleckchem.com/products/ABT-263.html were also similar, with BUCIS 03 having a slightly broader reference range than BUCIS 09 (BUCIS 03: 4.13–19.18 mg/L; BUCIS 09: 5.19–18.87 mg/L). In terms of the κ:λ ratio, the mAb assay had a similar range to Freelite™ (mAb
assay: 0.40–1.59; Freelite™: 0.58–1.76). Chlormezanone 1000 consecutive serum samples, selected as they arrived in the CIS for routine serum FLC analysis, were analysed using the mAb assay and Freelite™ (Fig. 4). Overall, each anti-FLC mAb showed good or excellent Spearman correlations with Freelite™: anti-κ BUCIS 01 (R2 = 0.79, 95% CI 0.76–0.81), anti-κ BUCIS 04 (R2 = 0.92, 95% CI 0.91–0.93), anti-λ BUCIS 03 (R2 = 0.87, 95% CI 0.85–0.88) and anti-λ BUCIS 09 (R2 = 0.85, 95% CI 0.84–0.87). Compared to each other, BUCIS 01 and BUCIS 04 mAbs provided a good correlation for κ FLC (R2 = 0.78, 95% CI 0.76–0.80) and BUCIS 03 and BUCIS 09 mAbs provided an excellent correlation for λ FLC (R2 = 0.97, 95% CI 0.97–0.98). In terms of the κ:λ ratio ( Fig. 5), both Freelite™ and the mAb assay demonstrated a good correlation (R2 = 0.85, 95% CI 0.83–0.86). Individual results from each assay were then compared to identify any discrepancies between the mAb assay and Freelite™. For this initial clinical validation of the mAb assay, the mean κ FLC results generated by BUCIS 01 and BUCIS 04 mAbs were used, and the λ FLC results obtained by BUCIS 03 and BUCIS 09 mAbs were used.