benefits over prior art use of polyclonal antibodies. The prior art "polyclonal" antibodies are said to
comprise "a population of different antibodies having varying specificities and affinities which
antibodies can be directed against different sites [or] determinants on the antigen (I)." Engvall
Application 06/539,754, specification, p. 5, lines 7-11. The use of monoclonals is said to allow more
accurate analysis and analytical procedures which have not been possible before. Engvall Application
06/539,754, specification, p. 5, lines 11-15. In the preferred embodiment, monoclonal antibodies (II)
and (III) react with structurally different determinants on the antigen. The advantage of this
embodiment is said to be that it permits the simultaneous addition of both monoclonal antibodies to
the antigen-containing solution. This eliminates an incubation and washing which is necessary with
prior art procedures using polyclonal antibodies. Engvall Application 06/539,754, specification, p. 5,
lines 18-25. Simultaneous addition of the two monoclonal antibodies is the preferred technique for
practicing the embodiment. Engvall Application 06/539,754, specification, p. 5, lines 25-28. In
discussing this preferred embodiment, neither affinity nor the affinity constant of the monoclonal
antibodies is not mentioned.
In another disclosed embodiment, the antibodies react with determinants which are equal or
structurally the same. In this embodiment the antibodies are reacted with the antigen sequentially.
The antigen first is reacted with the bound antibody and subsequently reacted with the unbound and
labeled antibody. Engvall Application 06/539,754, specification, p. 6, lines 1-13. In describing this
specific embodiment neither affinity nor the affinity constant is mentioned.
With respect to the monoclonal antibodies useful in the invention, the specification indicates
that the preparation of monoclonals is described in the prior art. The referenced prior art includes
the seminal work of Köhler and Milstein. Engvall Application 06/539,754, specification, p. 6, lines
14-21. The specification also describes a general method for preparation of monoclonal antibodies.
Engvall Application 06/539,754, specification, p. 6, line 22 - p. 7, line 10. The monoclonals may be
bound to a carrier and labeled according to prior art techniques. Engvall Application 06/539,754,
specification, p. 7, line 11 - p. 8, line 29. In describing the preparation of monoclonal antibodies, the
affinity constant is not mentioned.
The specification next presents three examples which are said to further illustrate the
invention. Engvall Application 06/539,754, specification, p. 9, lines 5-7. Examples 1 and 2 relate
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