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BMC Microbiol. 2015 Jul 3;15:133. doi: 10.1186/s12866-015-0461-1.

Differences in affinity of monoclonal and naturally acquired polyclonal antibodies against Plasmodium falciparum merozoite antigens.

Author information

1
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77, Stockholm, Sweden.
2
Department of Biochemistry, La Trobe University, Vic, 3086, Australia.
3
The Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria, Australia.
4
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
5
Papua New Guinea Institute of Medical Research, Goroka, 441, Papua New Guinea.
6
Swiss Tropical and Public Health Institute, Basel, Switzerland.
7
Institute for Glycomics, Griffith University, Queensland, Australia.
8
Department of Biochemistry, Makerere University, Kampala, Uganda.
9
Habib Medical School, IUIU, Kampala, Uganda.
10
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77, Stockholm, Sweden. kristina.persson@ki.se.
11
Department of Laboratory Medicine, Lund University, 22185, Lund, Sweden. kristina.persson@ki.se.

Abstract

BACKGROUND:

Malaria is a major global cause of deaths and a vaccine is urgently needed.

RESULTS:

We have employed the P. falciparum merozoite antigens MSP2-3D7/FC27 and AMA1, used them in ELISA, and coupled them in different ways using surface plasmon resonance (SPR) and estimated affinity (measured as kd) of monoclonal as well as naturally-acquired polyclonal antibodies in human plasma. There were major differences in kd depending on how the antigens were immobilized and where the His-tag was placed. For AMA1 we could see correlations with invasion inhibition. Using different immobilizations of proteins in SPR, we could see only moderate correlations with levels of antibodies in ELISA, indicating that in ELISA the proteins were not uniformly bound and that antibodies with many specificities exist in natural immunisation. The correlations between ELISA and SPR were enhanced when only parasite positive samples were included, which may indicate that high affinity antibodies are difficult to maintain over long periods of time. We found higher kd values for MSP2 (indicating lower affinity) compared to AMA1, which might be partly explained by MSP2 being an intrinsically disordered protein, while AMA1 is globular.

CONCLUSIONS:

For future vaccine studies and for understanding immunity, it is important to consider how to present proteins to the immune system to achieve highest antibody affinities.

PMID:
26149471
PMCID:
PMC4491891
DOI:
10.1186/s12866-015-0461-1
[Indexed for MEDLINE]
Free PMC Article

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