How to Calculate 4-Parameter Sigmoidal Curve

A four-parameter logistic (4PL) is best used when analyzing a sigmoidal, or S-shaped curve, a low-response curve or a high-response curve. If the curve is not symmetrical then it is better to use a five-parameter logistic (5PL). Create a plot of the concentration against the response applying the linear section of the response curve. This method is typically used to measure the results of enzyme-linked immunosorbent assays (ELISA).

Instructions

    • 1

      Write down the formula as follows:

      y = d + (a -- d)/ [1 + (x/c)b]

    • 2

      Find the value of a. This is the response value at zero concentration, or the minimum asymptote.

    • 3

      Find the value of b. This is the steepness of the curve, or Hill Slope, or slope factor. The steepness value can either be positive or negative.

    • 4

      Find the value of c. This is the point in the curve where the curvature of concavity changes directions, also known as the mid-range point.

    • 5

      Find the value of d. the response value for infinite concentration, or the maximum asymptote.

    • 6

      Determine x. This is the dilution. To find a base relative solution to run, follow the formula:

      Relative dilution = (actual sample / max dilution in series) * 100

    • 7

      Calculate for y. Once all the values are known, y can be calculated. These numbers are very accurate when determining the optical density for the sigmoidal shape.

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