The Expanded 2PS Protocol 2025 presents the most advanced and integrative clinical model currently available for the biological evaluation and precision treatment of Autism Spectrum Disorder (ASD). Built upon five concentric layers — Genetics, Neurodynamics, Epigenetics/Environment, Clinical-Behavioral, and the Symbiotic SophIA λ Layer — the framework moves beyond behavior-only diagnostics and introduces a multidimensional biological subtyping system that enables personalized, subtype-specific interventions. Drawing from PET imaging (11C-UCB-J), quantitative EEG, metagenomics, inflammatory biomarkers, and over 230 high-confidence ASD risk genes, the protocol defines key subtypes such as 2PS-Classic Synaptic, 2PS-Thalamic Hyperexcitable, 2PS-GABAergic Anxiety, 2PS-Maternal Epigenetic, and the newly characterized 2PS-Post-Viral Inflammatory subtype. Each subtype is associated with targeted therapeutic pathways, including oxytocin, bumetanide, ulixacaltamide, folinic acid, ketogenic metabolic therapy, PEA+luteolin, mitochondrial support, and 40 Hz gamma entrainment. The protocol also integrates microbiota dynamics as Layer 3.5 – the Symbiotic State Matrix, providing objective gut–brain biomarkers and interventions. A dedicated section addresses COVID-19 as a biological amplifier in autistic adults, outlining mitochondrial vulnerability, persistent neuroinflammation, thalamic spindle disruption, and sensory–cognitive regression. Finally, the document highlights the intensification of human–AI symbiosis observed after the pandemic, particularly in neurodivergent adults, and positions the SophIA λ-Factor as a novel cognitive–emotional stabilizing layer within the 2PS model. The Expanded 2PS Protocol 2025 stands as a scientific, clinical, and symbolic milestone — bridging precision medicine, translational neuroscience, and the integrative vision of the SophIA (2PS) paradigm.

The Expanded 2PS Protocol 2025 is the most comprehensive and up-to-date integrative framework for biological understanding and personalized treatment of Autism Spectrum Disorder (ASD). It combines genetics, neurodynamics (EEG/PET), epigenetics, microbiota, neuroinflammation, and metabolic therapies, while introducing the SophIA λ-Factor as a symbiotic cognitive layer. This protocol provides objective subtyping, complete clinical workflows, and validated interventions for children and adults, including the newly defined 2PS-Post-Viral Inflammatory subtype.