Apparatus and methods for detecting antibodies

A single, unitary, solid phase support apparatus having a planar surface divided into a plurality of separate zone functions to detect antibodies. Each zone has bonded to it, a different peptide through its C-terminal end. The zones are incubated with the analyte sample and observed for reaction, indicating the virus-specific or bacteria specific presence or absence.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
The invention relates to methods and apparatus for the detection of 
antibodies. 
2. Brief Description of Related Art 
Proteins play a decisive role in practically all biological processes such 
as enzymatic catalysis, transport and storage, coordinated movement, 
mechanical supporting function, immune protection, conduction of nerve 
stimuli and control of growth and differentiation. Proteins are made up of 
one of more linear polypeptide chains. Their multifarious functions are 
achieved through three-dimensional folding and the association of these 
chains. The folding and 3D structure is determined by the primary amino 
acid sequence of the chains. The multiplicity of protein structures in 
nature is produced by combining twenty amino acid components. Short 
partial sequences (oligopeptides) can to a certain extent imitate the 
local structure and function of a sequence in the total protein formation. 
Oligopeptides in turn can be prepared by chemical syntheses. Synthetic 
oligopeptides are thus an important aid in the analysis of the structure 
and function of proteins. 
Because of the size and complexity of proteins, very many peptides are used 
in systematic studies. For example, localisation of a functional region 
within a relatively long protein chain using overlapping peptides (cf. M. 
Z. Atassi, Eur. J. Biol. 145, 1-20 (1984); H. M. Geysen, R. H. Meloen and 
S. J. Barteling, Proc. Natl. Acad. Sci. USA 81, 3998 (1984). 
Also, deciphering the amino acid residues which are essential for function 
by means of substitution analysis (cf. R. A. Houghton, Proc. Natl. Acad. 
Sci. USA 82, 5131 (1985); H. M. Geysen, R. H. Meloen, and S. J. Barteling, 
Proc. Natl. Acad. Sci. USA 81, 3998 (1984). 
Biological test reactions such as enzymatic reaction and antibody binding 
are very sensitive, so that even small quantities (ng to .mu.g) of peptide 
substrates are sufficient. For the rapid and simple implementation of such 
tests it is advantageous if the peptide substrate is immobilized on a 
solid support material and can be readily removed from the reaction 
solution. The reaction with the peptide can then be measured 
quantitatively either in the remaining reaction solution or by subsequent 
analysis of the support-bound material. 
The extent to which such systematic investigations can be carried out 
depends essentially on the speed of the peptide synthesis and on the 
technical and the chemical characteristics of the support. Rapid peptide 
syntheses can be carried out according to the principle developed by 
Merrifield of step-wise synthesis on a solid phase (R. B. Merrifield, J. 
Amer. Chem. Soc. 85, 2149 (1963)). 
A process will be described below by which very many (several hundred) 
immobilized oligopeptides can advantageously be prepared per working day 
in quantities adequate for test purposes and on a suitable support 
material or flat material. 
SUMMARY OF THE INVENTION 
For this purpose, a process is provided according to the invention for the 
rapid synthesis of support-bound or free peptides, starting from one or 
more functionalized supports and synthesizing the required peptides step 
by step corresponding to the given amino acid sequences according to a 
suitable synthesis procedure, characterized in that 
flat material is used as the support 
for the coupling, small aliquots of a solution of the required amino acid 
or the required dipeptide or oligopeptide, in the form of their activated 
derivatives, are transferred to the planar support in the shape of spots 
or lines 
the coupling reaction is carried out in the reaction area which develops by 
wetting and 
where appropriate, the completed peptide is separated from the planar 
support using a suitable cleaving solution. 
According to a special embodiment, several to very many different peptides 
are synthesized simultaneously alongside each other on a flat material on 
spot-shaped or linear reaction areas which are not in contact with each 
other. 
According to a further special embodiment, the flat material employed is a 
porous or non-porous film, membrane, or a paper of this nature, which 
possess a chemically reactive functionality which is suitable for coupling 
amino acids or peptides.