Miniaturized Growth Inhibition Assay to Assess the Anti-blood Stage Activity of Antibodies

  • Elizabeth H. Duncan
  • Elke S. Bergmann-Leitner
Part of the Methods in Molecular Biology book series (MIMB, volume 1325)


While no immune correlate for blood-stage specific immunity against Plasmodium falciparum malaria has been identified, there is strong evidence that antibodies directed to various malarial antigens play a crucial role. In an effort to evaluate the role of antibodies in inhibiting growth and/or invasion of erythrocytic stages of the malaria parasite it will be necessary to test large sample sets from Phase 2a/b trials as well as epidemiological studies. The major constraints for such analyses are (1) availability of sufficient sample quantities (especially from infants and small children) and (2) the throughput of standard growth inhibition assays. The method described here assesses growth- and invasion inhibition by measuring the metabolic activity and viability of the parasite (by using a parasite lactate dehydrogenase-specific substrate) in a 384-microtiter plate format. This culture method can be extended beyond the described detection system to accommodate other techniques commonly used for growth/invasion-inhibition.

Key words

Plasmodium Antibodies Growth inhibition High-throughput Functional assay 



The authors would like to thank Dr. Wolfgang Leitner for critical reading of the manuscript.

Disclaimer The opinions or assertions contained herein are the private views of the authors, and are not to be construed as official, or as reflecting true views of the Department of the Army or the Department of Defense.


  1. 1.
    Dent AE, Bergmann-Leitner ES, Wilson DW et al (2008) Antibody-mediated growth inhibition of Plasmodium falciparum: relationship to age and protection from parasitemia in Kenyan children and adults. PLoS One 3(10), e3557Google Scholar
  2. 2.
    John CC, Moormann AM, Pregibon DC et al (2005) Correlation of high levels of antibodies to multiple pre-erythrocytic Plasmodium falciparum antigens and protection from infection. Am J Trop Med Hyg 73(1):222–228Google Scholar
  3. 3.
    John CC, O’Donnell, Sumba PD et al (2004) Evidence that invasion-inhibitory antibodies specific for the 19-kDa fragment of the merozoite surface protein-1 (MSP-119) can play a protective role against Blood-stage Plasmodium falciparum infection in individuals in a malaria endemic area of Africa. J Immunol 173:666–672Google Scholar
  4. 4.
    Thera MA, Doumbo OK, Coulibaly B et al (2010) Safety and immunogenicity of an AMA1 malaria vaccine in Malian children: results of a phase 1 randomized controlled trial. PLoS One 5(2), e9041Google Scholar
  5. 5.
    Bergmann-Leitner ES, Duncan EH, Mullen GE et al (2006) Critical evaluation of different methods for measuring the functional activity of antibodies against malaria blood stage antigens. Am J Trop Med Hyg 75(3):437–442Google Scholar
  6. 6.
    Ringwald P, Meche FS, Bickii J et al (1999) In vitro culture and drug sensitivity assay of Plasmodium falciparum with nonserum substitute and acute-phase sera. J Clin Microbiol 37(3):700–705Google Scholar
  7. 7.
    Bergmann-Leitner ES, Duncan EH, Burge JR et al (2008) Short report: miniaturization of a high-throughput pLDH-based Plasmodium falciparum growth inhibition assay for small volume samples from preclinical and clinical vaccine trials. Am J Trop Med Hyg 78(3):468–471Google Scholar
  8. 8.
    Bergmann-Leitner ES, Leitner WW (2008) Antibody purification. In: Biological research methodology—a handbook. Taylor&Francis Group Publishers/CRC Press ISBN 9780849376160Google Scholar
  9. 9.
    Bergmann-Leitner ES, Mease RM, Duncan EH et al (2008) Evaluation of immunoglobulin purification methods and their impact on quality and yield of antigen-specific antibodies. Malar J 7(1):129Google Scholar
  10. 10.
    Haynes JD, Moch JK (2002) Automated synchronization of Plasmodium falciparum parasites by culture in a temperature-cycling incubator. Methods Mol Med 72:489–497Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Elizabeth H. Duncan
    • 1
  • Elke S. Bergmann-Leitner
    • 1
  1. 1.Malaria Vaccine BranchWalter Reed Army Institute of ResearchSilver SpringUSA

Personalised recommendations