Over the past several decades, biomarkers have become indispensable tools in human biology, allowing researchers to investigate biocultural and evolutionary questions and to generate basic data on health and well-being. Human biologists are central players in biomarker methods development, pioneering the creation of "field friendly," minimally invasively collected biomarker approaches, as well as leading the innovative use of biomarkers, most notably to map the complex pathways from social and environmental conditions to altered physiology to health effects. This special issue, Part 2 of a collection focused on minimally invasively collected biomarkers, is comprised of nine papers that jointly contribute to driving the science of biomarker methods development and application. This compilation of papers engages with topics such as biological normalcy, ecoimmunology, and the ethics of working with vulnerable and underserved populations. It also focuses attention on research areas at present not emphasized in human biology such as bone turnover markers as a window onto osteoporosis and osteoarthritis and the use of cancer-related biomarkers in population screening and epidemiology. Taken together, these papers help draw the roadmap for future biomarker work, emphasizing: (1) the need for systematic and transparent approaches to assay validation, with open access publication; (2) simultaneous measurement of multiple biomarkers and expanded use of instrument platforms that increase the range of physiological, genomics, and omics markers available to researchers; and, (3) increased attention to ethics and researcher responsibilities, encouraging a mindset that recognizes our obligation to provide benefits to individuals and communities and to help redress past abuses.