The recently proposed multi-chirp waveform, affine frequency division multiplexing (AFDM), is regarded as a prospective candidate for integrated sensing and communication (ISAC) due to its robust performance in high-mobility scenarios and full diversity achievement in doubly dispersive channels. However, the insufficient Doppler resolution caused by limited transmission duration can reduce the accuracy of parameter estimation. In this paper, we propose a new off-grid target parameter estimation scheme to jointly estimate the range and velocity of the targets for AFDM-ISAC system, where the off-grid Doppler components are incorporated to enhance estimation accuracy. Specifically, we form the sensing model as an off-grid sparse signal recovery problem relying on the virtual delay and Doppler grids defined in the discrete affine Fourier (DAF) domain, where the off-grid components are regarded as hyper-parameters for estimation. We also employ the expectation-maximization (EM) technique via a sparse Bayesian learning (SBL) framework to update hyper-parameters iteratively. Simulation results indicate that our proposed off-grid algorithm outperforms existing algorithms in sensing performance and is highly robust to the AFDM-ISAC high-mobility scenario.