We prove an analogue of Alon’s spectral gap conjecture for random bipartite, biregular graphs. We use the Ihara–Bass formula to connect the non-backtracking spectrum to that of the adjacency matrix, employing the moment method to show there exists a spectral gap for the non-backtracking matrix. A by-product of our main theorem is that random rectangular zero-one matrices with fixed row and column sums are full rank with high probability. Finally, we illustrate applications to community detection, coding theory, and deterministic matrix completion.