Medical Microbiology and Immunology

, Volume 165, Issue 4, pp 271–288 | Cite as

Immunochemical properties of oligosaccharide-protein conjugates withKlebsiella-K2 specificity

I. Specificity and crossreactivity of anti-conjugate versus anti-bacterial antibodies
  • Hildegard Geyer
  • Stephan Stirm
  • Karl Himmelspach


A series of repeating unit oligomers
$$\begin{gathered} \alpha - GlcUA \hfill \\ 1 \downarrow 3 \hfill \\ [ \to 4) - \beta - Man - (1 \to 4) - \alpha - Glc - (1 \to 3) - \beta - Glc - (1 \to ]_n , 1 \leqslant n \leqslant 7 , \hfill \\ \end{gathered} $$
was prepared by depolymerization ofKlebsiella pneumoniae B5055 (01∶K2) capsular polysaccharide (K2-PS) as catalyzed by a bacteriophage-associated glycanase. The monomeric repeating unit [tetrasaccharide, (TS)] and its di- and trimer [octa- and dodecasaccharide (OS and DS)] were conjugated to edestin via reductive aminophenylation and azo coupling at the reducing sugar end group. Rabbit anti-conjugate and anti-bacterial antibodies raised in rabbits were compared with respect to specificity and crossreactivity towards the oligomers of the various molecular sizes, towards parent PS, and towards whole bacteria. Antibodies able to bind specifically to the PS and the bacteria were elicited by all three conjugates. However, the anti-TS conjugate antibodies, in contrast to those obtained with OS and DS conjugate, proved to be practically unable to effect bacterial agglutination. Correspondingly, the TS played an exceptional role in binding to anti-bacterial antibodies. In contrast to the OS and DS it could not fully inhibit precipitation of these antibodies with the bacterial PS. Moreover, the inhibition of the binding of the PS to antibacterial antibodies produced by TS was about 50-fold weaker than that produced by OS, DS, and higher members of the series, all of which were about equally potent inhibitors (on a molar basis). The results show that two repeating units are the minimum requirement for a substantial representation of the PS's serologic specificity. The exceptional behavior of the TS correlates with its lack of theβ-Glc-(1–4)-Man linkage present in all higher members of the series.


Oligomer Capsular Polysaccharide Molar Basis Tetrasaccharide Conjugate Antibody 
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phosphate-buffered saline


plaque-forming units




Klebsiella pneumoniae B5055 (01∶K2) capsular polysaccharide


tetrasaccharide (=monomeric repeating unit)


octasaccharide (=dimeric repeating unit)


dodecasaccharide (=trimeric repeating unit)


sodium cyanoborohydride


human γ-globulin


Freund's complete adjuvant





TS125I, PS125I

iodine-125-labelled TS or K2-PS, respectively


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Copyright information

© Springer-Verlag 1979

Authors and Affiliations

  • Hildegard Geyer
    • 1
  • Stephan Stirm
    • 1
  • Karl Himmelspach
    • 1
  1. 1.Max-Planck-Institut für ImmunbiologieFreiburg/Br.Federal Republic of Germany

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