| Summary: | Polysaccharides with molecular weights ranging from 1.75 × 10<sup>3</sup> to 1.14 × 10<sup>4</sup> g/mol were obtained from the fruit bodies of <i>Ganoderma lucidum</i>. The multiple fingerprints and macrophage immunostimulatory activity of these fractions were analyzed as well as the fingerprint–activity relationship. The correlation analysis of molecular weight and immune activity demonstrated that polysaccharides with molecular weights of 4.27 × 10<sup>3</sup>~5.27 × 10<sup>3</sup> and 1 × 10<sup>4</sup>~1.14 × 10<sup>4</sup> g/mol were the main active fractions. Moreover, the results showed that galactose, mannose, and glucuronic acid were positively related to immunostimulatory activity. Additionally, partial least-squares regression and grey correlation degree analyses indicated that three peaks (P2, P3, P8) in the oligosaccharide fragment fingerprint significantly affected the immune activity of the polysaccharides. Hence, these ingredients associated with activity could be considered as markers to assess <i>Ganoderma lucidum</i> polysaccharides and their related products, and the study also provides a reference for research on the spectrum–effect relationship of polysaccharides in the future.
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