Phagotopes derived by antibody screening of phage-displayed random peptide libraries vary in immunoreactivity: Studies using an exemplary monoclonal antibody, CII-C1, to type II collagen

Antibody screening of phage-displayed random peptide libraries to identify mimotopes of conformational epitopes is promising. However, because interpretations can be difficult, an exemplary system has been used in the present study to investigate whether variation in the peptide sequences of selected phagotopes corresponded with variation in immunoreactivity. The phagotopes, derived using a well-characterized monoclonal antibody, CII-C1, to a known conformational epitope on type II collagen, C1, were tested by direct and inhibition ELISA for reactivity with CII-C1. A multiple sequence alignment algorithm, PILEUP, was used to sort the peptides expressed by the phagotopes into clusters. A model was prepared of the C1 epitope on type II collagen. The 12 selected phagotopes reacted with CII-C1 by both direct ELISA (titres from < 100-11 200) and inhibition ELISA (20-100% inhibition); the reactivity varied according to the peptide sequence and assay format. The differences in reactivity between the phagotopes were mostly in accord with the alignment, by PILEUP, of the peptide sequences. The finding that the phagotopes functionally mimicked the C1 epitope on collagen was validated in that amino acids RRL at the amino terminal of many of the peptides were topographically demonstrable on the model of the C1 epitope. Notably, one phagotope that expressed the widely divergent peptide C-IAPKRHNSA-C also mimicked the C1 epitope, as judged by reactivity in each of the assays used: these included cross-inhibition of CII-C1 reactivity with each of the other phagotopes and inhibition by a synthetic peptide corresponding to that expressed by the most frequently selected phagotope, RRLPFGSQM. Thus, it has been demonstrated that multiple phage-displayed peptides can mimic the same epitope and that observed immunoreactivity of selected phagotopes with the selecting mAb can depend on the primary sequence of the expressed peptide and also on the assay format used.

Henny? Beitman with a bicycle; Dielkirchen, Falkenstein, Germany Portraits Women

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Mr. and Mrs. Fuerst Berlin, Hans Frankenbach, Wronker Slogau? and Lilienthal Berlin seated at an outdoor table; Brambach, Germany Portraits Goups

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Jewish refugees from Warsaw waiting at the border train station for transport to Germany Portraits; Groups

Weltkrieg 1914/18 Aus Warschau gefluechtete Juden, die auf einem Grenzbahnhof auf Abtransport nach Deutschland warten

Exterior view of Schoenbeck residence; Stadtoldendorf, Germany.

Photo owned by Annette Rochaix, Belmont Switz. Deposited: Archiv Cantonales, Remens, Switz. Copy by T. Krakauer

Gottschalk children at swim club forming a pyramid; Hannover, Germany.

Seated on grass Kurt Godshaw; left to right Walter, Freddy, Ursula; seated on ball Hal Godshaw

Freddy Godshaw (Gottschalk) sailing with his friend Peter Zander at a camp shortly after Freddy left Germany; England.

Left: Freddy Godshaw (Gottschalk); right: Peter Zander

Gottschalk children at swim club forming a pyramid; Hannover, Germany.

Seated on grass Kurt Godshaw; left to right Walter, Freddy, Ursula; seated on ball Hal Godshaw

Kurt Godshaw (Gottschalk) as a teenager lying in a meadow; Hannover, Germany.

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Studio portrait of Walter Godshaw (Gottschalk) as a teenager; Hannover, Germany.

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Wedding portrait of Hans Krakauer and Elizabeth Godshaw (Gottschalk); Hannover, Germany. Portraits Couples

In the apartment on Ruemkorffstrasse that the Gottschalks occupied after being forced to sell their house.

Elizabeth Krakauer nee Gottschalk seated in a Oberprima (high school) chemistry laboratory; Sophienschule; Hannover, Germany.

This picture was taken during her last year of high school. The chemistry teacher, Professor Schmigielski was one of Elizabeth's favorite teachers.

Portrait of Elizabeth Krakauer nee Gottschalk with pigtails; Hannover, Germany.

This portrait shows Elizabeth with pig tails. In a defiant mood, she cut them off.

Exterior view of Gottschalk residence; Seelhorst Strasse 27, Hannover, Germany.

Fig. 13 in family portrait

Studio portrait of Josef Molling with unidentified man; Heidelberg, Germany.

right: Joseph Molling sitting; left: unknown man standing