Acinetobacter baumannii is a high priority pathogen, rapidly acquiring multidrug resistance. Limited temporal resolution at the proteomic and transcriptomic levels has constrained our understanding of real-time resistance acquisition, thus hindering the development of strategies to combat resistance. The study aimed to unveil the resistance acquisition processes. A total of 45 clinical A. baumannii strains from three geographic locations were exposed to sub-inhibitory levels of carbapenem, aminoglycoside and β-lactam antibiotics for 72 hours. Kinetic resistance acquisition was simultaneously monitored by Western blot for expression of resistance proteins (β-lactamases, efflux pumps, porins) and quantitative PCR (qPCR) for mRNA using custom TaqMan probes with highest temporal resolution (13 timepoints at 6h intervals). Western blot analyses identified novel biphasic induction of resistance proteins: early phase (0–18h) with 3.4-fold induction of OXA-23, and late phase (24–72h) with 7.8-fold induction of NDM-1. qPCR results found mRNA levels reached their peak 4–8 hours prior to peak protein expression, suggesting robust post-transcriptional regulation. New discovery: AdeB efflux pump protein showed 12.6-fold increase with only 2.1-fold mRNA increase, suggesting extreme translational amplification with protein/mRNA ratio of 6.1. Machine learning algorithm categorized response into three distinct patterns with profound clinical implications: early responders (89% treatment failure); late responders (45% failure); constitutive expressors (67% failure). Thus, integrative analyses identified novel temporal dynamics of resistance development, key 4–8 hour windows for intervention before resistance is irreversible. Post-transcriptional regulation is much more important than previously thought, and may be exploited for the development of new combination therapies and individualised anti-microbial therapies.