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Table 2 Kinetic parameters obtained from the optimization of the Peptidomatrix conditions for peptide MXR P2 and the serum from rabbit # 64850 (see Figure 2) (± SD, n = 3).

From: Competition between bound and free peptides in an ELISA-based procedure that assays peptides derived from protein digests

A : increasing attached peptide at four fixed levels of added antibody

 

1 stAB Concentration 0.0005 serum/PBS

1 stAB Concentration 0.001 serum/PBS

1 stAB Concentration 0.005 serum/PBS

1 stAB Concentration 0.01 serum/PBS

Kd (μg/ml)

0.49 ± 0.25

0.89 ± 0.56

0.49 ± 0.097

0.24 ± 0.06

Amax (OD units)

0.076 ± 0.01

0.12 ± 0.01

0.22 ± 0.012

0.35 ± 0.024

D (OD units)

0.29 ± 0.008

0.3 ± 0.015

0.29 ± 0.009

0.3 ± 0.02

B: increasing added antibody at three fixed levels of attached peptide

 

Attached peptide 0.25 μg/ml

Attached peptide 0.5 μg/ml

Attached peptide 1 μg/ml

 

Kd (serum dilutions)

0.003 ± 0.0007

0.003 ± 0.001

0.001 ± 0.0002

 

Amax (OD units)

0.35 ± 0.023

0.56 ± 0.06

0.49 ± 0.02

 

D (OD units)

0.38 ± 0.01

0.38 ± 0.02

0.38 ± 0.01

 

C: Competition assay – Kd determined when increasing added free peptide at two fixed levels of attached peptide

 

Attached peptide 0.25 μg/ml

Attached peptide 0.5 μg/ml

  

Kd (μg/ml)

0.14 ± 0.03

0.97 ± 0.15

  

Amax (OD units)

0.29 ± 0.019

0.37 ± 0.016

  

D (OD units)

0.39 ± 0.01

0.35 ± 0.01

 Â