## Description of ELISAeq Computer Program

This Computer program is written in GW-BASIC we have named ELISAeq. The main function of this program is to derive the linear \& hyperbolic interpolation formula using the known standard and to determine the Ag/Ab (Antibody or antigen) concentration of unknown samples. The ELISAeq have three main sections I) Input Section (Lines 270--2000) II) Subroutine section (lines 2400--3800) and III) Calculation \& Result section (lines 2010--2400). The detail description of each section is as below.

Input Section: This section of the program allow the user to input information. Lines 270--720 allow to feed the optical density (OD) data from ELISA plate. The OD data can be feed directly from microplate reader by interfacing its output port to the serial port of microcomputer using RS 232-C interface, or by using keyboard or file. Lines 820--1280 allow to display the OD data in $8\times 12$ format on screen, to modify the OD data, to store this OD data for future use, and also allow to print the OD data in $8\times 12$ on printer. Lines 1330--2000 allow to feed the dilution template, which is the information about known standard such as the rows containing the standard samples, the concentration of these known standards and whether the samples are in singlet, duplicates, triplicates etc. This dilution template can be feed by using keyboard or by using the file, and can be stored for future use.

Subroutine Section: This section consist three subroutines I) Linear regression II) Hyperbolic regression and III) Error trapping. Linear regression subroutine (lines 3340--3540) derive a linear interpolation formula using the linear regression and least square curve fitting method . The hyperbolic regression subroutine (lines 3550--3830) derive a hyperbolic interpolation formula using the hyperbolic regression method. Error trapping (lines 3840--3930) subroutine handle the errors occurs, when the program is running.

Calculation & Result section This section perform the required calculation by utilizing the subroutine section. Lines 2020--2410 allow to calculate the average value of OD if the samples on the plate are in duplicate, triplicate etc, and also allow to display \& print the average value. The program utilized the standard data to determine the Sl-range (semilogarithmic linear range) and derive a linear interpolation formula for this range by using linear regression and least square curve-fitting method. The hyperbolic interpolation formula was also derived by using hyperbolic regression method from standard data. The OD of unknown samples was checked and Ab/Ag concentration determined by linear interpolation formula whose OD is in the Sl-range and by hyperbolic interpolation formula whose OD is beyond the Sl-range. It also calculate the signal, capacity and flatness of hyperbolic curve, which play a vital role in ELISA optimization. The program allow to display and print the results.

Files contained in this package:
FILES DESCRIPTION
ELISAEQ.BAS SOURCE CODE OF ELISAEQ, IN GW-BASIC
OD.DAT EXAMPLE FILE CONTAIN OPTICAL DENSITY DATA
DT.DAT EXAMPLE FILE CONTAIN DILUTION TEMPLATE DATA
PAPER.TEX MANUSCRIPT IN TEX FORMAT (NON-DOCUMENT) FILE
elisaeq.tar All files in tar format
elisaeq.tar.uue uuencoded elisaeq.tar file

LICENCE:

This program remains the property of the Institute of Microbial Technology, Chandigarh, INDIA. This program may be freely used by anybody subject to the following conditions:

1. The authors nor the Institute of Microbial Technology Chandigarh assume any responsibility for any losses or damage that may be caused by the use or misuse of the accompanying software.

2. The authors nor the Institute of Microbial Technology Chandigarh give any warranty with regards to the software being able to function on any computer.

3. The accompanying software may not be copied nor distributed with any modifications, and this document file MUST be included with all copies.

4. No fee may be charged for the copying and/or distribution of the accompanying software.

5. Users must agree to accept any risk as a condition of the free use of the accompanying software.

Any suggestion, bug report or comment will be greatly appreciated. Please send them to:
               G P S Raghava, Scientist

Co-ordinator, Bioinformatics Center

Institute of Microbial Technology,

Sector 39A, Chandigarh, India.