This web server allow to compute the endpoint titer of antibodies and concentration of antbody/antgen concentration from ELISA data. This server is based on program ELISAeq which is freely available from Home Page Raghava or Archive of Journal of Immunological methods. Input data required and terms used in web server are described below.
Input Data
Ab/Ag Concentration: In order to calculate the concentration of Ab or Ag one should include known standard on plate. The concentration of these known standard and corresponding optical density (OD) is used as input for the web page. This input is to drive the equation which would serve to estimate the oncentration of unknown samples.
In order to calculate the Ab/Ag concentration one should incorporate the concentration and OD of known standards. The CGI script which is written in PERL, will fit this data using graphical method and will generate another web page which include the java-script. This would allow the user to compute the concentration of unknown samples.
Alernatively, one can also calculate the concentration of Ab or Ag of unkown sample even when concentration of standards is not known. In this case one should put the known standard in serial dilutions. The dilution factor and OD of standard can be used as input. In this case, CGI script will fit the dilution fold and OD and generate a web page for calculating dilution fold of unknown samples.
Endpoint Titer: In order to compute the endpoint antibody titer of test sample, ODs of serial dilutions of known samples and that of control (i.e. backgroung OD of test sample in the absance of antigen) should be used as input. User should feed concentration and corresponding OD of in column 'Standard or Known samples' and concentration and corresponding OD of control samples in column 'Control Data' . The concentration and corresponding OD of known standards and control sample will be used as input for calculating endpoint titer.
The CGI script will fit the known standard using graphical method and control samples using semi-lograthmic linear curve. It will also findout the intersection or nearest point of two curve. The concentration at this point will be shown as endpoint titer.
Alternatively, one can calculate the endpoint titer in terms of dilution. In this case user should provide the dilution factor and corresponding OD of these serial dilution of both standard and control data.
These are the samples whose concentration is known. These known samples
should be included on ELISA plate in order to calculate the concentration
of unknown samples. The user should input the Ab or Ag concentration and
corresponding OD in respective columns.:
1 (a) Ab/Ag Conc.(S): User should feed the concentrations
of standard or known samples in this column; or
1(b) DF(S): In case concentration of standard
is not available then the user can put the standards by making serail dilutions
using a dilution factor (e.g., Log5 dilutions).
2 OD(S): User should feed the OD of serial dilutions
of test/unknown samples in this column corresponding to concentration in
column Ab/Ag Conc.(S).
Control Data:
These are the standard samples whose run in absance of any interaction
(antbody in absance of antigen). These samples should be included on ELISA
plate in order to calculate the endpoint titer.
1 (a) Ab/Ag Conc.(C): User should feed the concentration
of antibody in absance of antigen of standard in this column, or
1 (b) DF(C): In case concentration of standard
is not known then the user can put the standards in by making serail dilutions
using a dilution factor (e.g, log5 dilutions).
2 OD(C): User should feed the OD of test/unknown
samples in this column corresponding to concentration in column Ab/Ag
Conc.(C). In case concentration of standard sample is not known then
the OD for serail dilutions should be fed in this column.