Replace multiple characters in one replace call

Use the OR operator (|):

var str="#this #is__ __#a test###__";
str.replace(/#|_/g,''); // result: "this is a test"

You could also use a character class:

str.replace(/[#_]/g,'');

Fiddle

If you want to replace the hash with one thing and the underscore with another, then you will just have to chain. However, you could add a prototype:

String.prototype.allReplace = function(obj) {
    var retStr = this;
    for (var x in obj) {
        retStr = retStr.replace(new RegExp(x, 'g'), obj[x]);
    }
    return retStr;
};

console.log('aabbaabbcc'.allReplace({'a': 'h', 'b': 'o'}));
// console.log 'hhoohhoocc';

Why not chain, though? I see nothing wrong with that.