1Analysis of the spatial sub-cellular distribution of proteins is of vital importance 2 to fully understand context specic protein function. Some proteins can be found 3 with a single location within a cell, but up to half of proteins may reside in multiple 4 locations, can dynamically re-localise, or reside within an unknown functional com-5 partment. These considerations lead to uncertainty in associating a protein to a single 6 location. Currently, mass spectrometry (MS) based spatial proteomics relies on super-7 vised machine learning algorithms to assign proteins to sub-cellular locations based on 8 common gradient proles. However, such methods fail to quantify uncertainty associ-9 ated with sub-cellular class assignment. Here we reformulate the framework on which 10 we perform statistical analysis. We propose a Bayesian generative classier based on 11 Gaussian mixture models to assign proteins probabilistically to sub-cellular niches, thus 12 proteins have a probability distribution over sub-cellular locations, with Bayesian com-13 putation performed using the expectation-maximisation (EM) algorithm, as well as 14 Markov-chain Monte-Carlo (MCMC). Our methodology allows proteome-wide uncer-15 tainty quantication, thus adding a further layer to the analysis of spatial proteomics. 16 Our framework is exible, allowing many dierent systems to be analysed and reveals 17 new modelling opportunities for spatial proteomics. We nd our methods perform 18 competitively with current state-of-the art machine learning methods, whilst simulta-19 neously providing more information. We highlight several examples where classication 20 based on the support vector machine is unable to make any conclusions, while uncer-21 tainty quantication using our approach provides biologically intriguing results. To our 22 knowledge this is the rst Bayesian model of MS-based spatial proteomics data. 23 * omc25@cam.ac.uk † lg390@cam.ac.uk Author summary 24Sub-cellular localisation of proteins provides insights into sub-cellular biological processes. 25 For a protein to carry out its intended function it must be localised to the correct sub-26 cellular environment, whether that be organelles, vesicles or any sub-cellular niche. Correct 27 sub-cellular localisation ensures the biochemical conditions for the protein to carry out its 28 molecular function are met, as well as being near its intended interaction partners. Therefore, 29 mis-localisation of proteins alters cell biochemistry and can disrupt, for example, signalling 30 pathways or inhibit the tracking of material around the cell. The sub-cellular distribution 31 of proteins is complicated by proteins that can reside in multiple micro-environments, or 32 those that move dynamically within the cell. Methods that predict protein sub-cellular 33 localisation often fail to quantify the uncertainty that arises from the complex and dynamic 34 nature of the sub-cellular environment. Here we present a Bayesian methodology to analyse 35 protein sub-cellular localisation. We ex...