Graduation Year


Document Type




Degree Name

Master of Arts (M.A.)

Degree Granting Department


Major Professor

Elizabeth Miller, Ph.D.

Committee Member

David Himmelgreen, Ph.D.

Committee Member

Lorena Madrigal, Ph.D.


breastmilk, immunity, immunoassay, life history, TGF-β2


Objectives: There are three aims for this thesis: the first is to develop a field and laboratory protocol for the storage and analysis of transforming growth factor–beta 2 (TGF-β2) in human breastmilk; second, to validate this protocol and the immunoassay used to assess this new method; and lastly, to explore the ramifications of biosimilar TGF-β2 across multiple milks on human health, growth, and immunity through the review of laboratory findings and previous literature.

Rational: Little anthropological research has been done on TGF-β2 in human milk. Anthropology as a discipline is well positioned to provide insight into TGF-β2, combining biocultural, evolutionary, and ecological approaches to holistically illustrate the effects this cytokine has on human immunity. This thesis provides an applied anthropological perspective and methodology on TGF-β2 in human milk.

Methods: A protocol was developed for a new method of drying breastmilk on polystyrene microplates. Samples were then reconstituted using reagent diluent with 1% BSA and assayed using a Human TGF-beta 2 DuoSet enzyme-linked immunosorbent assay (ELIZA) assay kit from R&D Systems. Other mammalian milks and infant formula samples were also dried and tested for TGF-β2 concentrations. Validity of the assay and TGF-β2 concentrations were then statistically measured using linear regression analysis and Bland-Altman plots.

Results: The results of the first objective in the development of a laboratory and field protocol for drying breastmilk on polystyrene plates for the extraction of TGF-β2 showed this method to hold promise for future application, but lacked statistical power in this study to confirm if this method is viable. The second objective of assay validation was unsuccessful, with the percent coefficient of variation for the intra-assay variation and inter-assay validation 38.28% and 17.70%, respectively indicating that this assay struggled to produce consistent and reliable results from the reconstituted samples. Results from the third objective suggest that biosimilar TGF-β2 in non-human milk can influence human growth and development, the extent of which, however, needs further study.

Conclusions: Given these findings, more work with TGF-β2 in milk is required. TGF-β2 is a cytokine which could reveal a great deal about the developmental origins of human immunity and how it is maintained and altered across our life course and therefore an area of biology worth further research.

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Biology Commons