Development of a second generation Inhibin B ELISA.
Kalra. Bhanu B; Kumar. Ajay A; Patel. Kinita K; Patel. Amita A; Khosravi. M J MJ
Key Findings
- A sandwich‑type ELISA can measure Inhibin B in 50 µL of serum or plasma in about 3.5 hours.
- The assay is highly specific: it does not react with related proteins such as follistatin‑315, Activins, AMH, FSH, or LH.
- Performance metrics (precision, linearity, limit of quantitation) are comparable or better than two commercial Inhibin B kits.
Practical Outcomes
- For DIY biohackers, this study doesn’t provide a new protocol, dosage, or health benefit. It simply offers a better laboratory tool for researchers who want to measure Inhibin B, which may eventually support studies on fertility or metabolic health, but there are no immediate actionable takeaways for personal use.
Summary
The paper describes a new lab test for measuring the hormone Inhibin B in blood. It shows the test is fast, accurate, and doesn’t get confused by other similar proteins, including follistatin‑315, but it doesn’t give any advice on how to use Inhibin B or follistatin‑315 for health or performance.
Abstract
Inhibins are heterodimeric protein hormones secreted by the granulosa cells of the ovary in the female and the Sertoli cells of the testis in the male. Published research studies have assessed Inhibin B levels in Sertoli cell function, ovarian reserve and granulosa cell tumors. A two-step sandwich-type enzymatic microplate assay to measure Inhibin B levels within 3.5h is reported, and sample pre-treatment is not required. The assay measures Inhibin B in 50 μL of serum or Li-Hep plasma sample against Inhibin B calibrators (10-1000 pg/mL). The highly characterized antibody pair used in the assay measures 100% Inhibin B and no response was detected above the sensitivity of the assay with Inhibin A, Activin A, Activin B, Activin AB, AMH, FSH, LH or Follistatin 315 at the concentrations tested. The second generation Inhibin B assay was compared against two commercially available assays using 60 male and 60 female samples, ranging in age from 20 to 50 years. The assay showed significant positive linear correlations to Oxford Brooks Innovation (OBI) and Diagnostics Systems Laboratories (DSL) assays (r=0.99; P<0.0001; and r=0.97; P<0.0001), respectively. Method comparison to OBI and DSL resulted in the following slope and intercept (Gen II=1.03 OBI-6.77 pg/mL and Gen II=1.57 DSL+11.29 pg/mL), respectively. Matched serum and Li-Hep plasma samples (n=120) showed a correlation coefficient of >0.99 and a slope of 0.97 with zero intercept. Total imprecision calculated on three samples and two controls over 40 runs, three replicates per run, using NCCLS EP5-A guidelines was 6.8% at 19.34 pg/mL, 4.4% at 76.03 pg/mL, 4.3% at 275.3 pg/mL, 5.4% at 99.88 pg/mL, and 5.7% at 363.9 pg/mL. The LoQ for the assay at 20% CV was 4.8 pg/mL. Dilution and spiking studies showed an average recovery of 90-110%. A highly specific, sensitive, simplified and reproducible microplate Inhibin B assay has been developed to measure Inhibin B in serum and Li-Hep plasma. The performance of the assay is ideal for investigation into the physiologic role of Inhibin B in both men and women.
Study Information
pubmed
2010
2010-08-21T00:00:00.000Z
10.1016/j.jim.2010.08.002
26
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